isoelectric focusing

Summary

Summary: Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point.

Top Publications

  1. ncbi Isoelectric focusing and two-dimensional gel electrophoresis
    David B Friedman
    Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee, USA
    Methods Enzymol 463:515-40. 2009
  2. pmc Peptide separations by on-line MudPIT compared to isoelectric focusing in an off-gel format: application to a membrane-enriched fraction from C2C12 mouse skeletal muscle cells
    Sarah Elschenbroich
    Ontario Cancer Institute, University Health Network, Canada
    J Proteome Res 8:4860-9. 2009
  3. ncbi Laboratory testing of individuals with severe alpha1-antitrypsin deficiency in three European centres
    M Miravitlles
    Fundació Clínic, Institut d Investigacions Biomediques August Pi i Sunyer, CIBER de Enfermedades Respiratorias, Barcelona, Spain
    Eur Respir J 35:960-8. 2010
  4. ncbi Kinetics of local and systemic isoforms of serum amyloid A in bovine mastitic milk
    S Jacobsen
    Department of Clinical Studies, Large Animal Surgery, Royal Veterinary and Agricultural University, Dyrlaegevej 48, DK 1870 Frederiksberg C, Copenhagen, Denmark
    Vet Immunol Immunopathol 104:21-31. 2005
  5. ncbi Serum amyloid A isoforms in serum and synovial fluid from spontaneously diseased dogs with joint diseases or other conditions
    Mads Kjelgaard-Hansen
    Central Laboratory, Department of Small Animal Clinical Sciences, Faculty of Life Sciences, University of Copenhagen, Groennegaardsvej 3, DK 1870 Frederiksberg, Denmark
    Vet Immunol Immunopathol 117:296-301. 2007
  6. ncbi Identification of distinct N-terminal truncated forms of prion protein in different Creutzfeldt-Jakob disease subtypes
    Gianluigi Zanusso
    Department of Neurological and Visual Sciences, Section of Neurology and Agricultural and Industrial Biotechnologies, University of Verona, 37134 Verona, Italy
    J Biol Chem 279:38936-42. 2004
  7. ncbi Current two-dimensional electrophoresis technology for proteomics
    Angelika Görg
    Department of Proteomics, Technische Universitat Munchen, Freising Weihenstephan, Germany
    Proteomics 4:3665-85. 2004
  8. pmc Animal and human multidrug-resistant, cephalosporin-resistant salmonella isolates expressing a plasmid-mediated CMY-2 AmpC beta-lactamase
    P L Winokur
    University of Iowa College of Medicine, Iowa City, Iowa, USA
    Antimicrob Agents Chemother 44:2777-83. 2000
  9. ncbi Quantitative membrane proteomics applying narrow range peptide isoelectric focusing for studies of small cell lung cancer resistance mechanisms
    Hanna Eriksson
    Karolinska Biomics Center, Karolinska University Hospital, Stockholm, Sweden
    Proteomics 8:3008-18. 2008
  10. ncbi The use of sigmoid pH gradients in free-flow isoelectric focusing of human endothelial cell proteins
    Xiao Ma
    Institute of Biomedicine, Pharmacology, Aarhus University, Aarhus, Denmark
    Electrophoresis 33:1349-55. 2012

Detail Information

Publications314 found, 100 shown here

  1. ncbi Isoelectric focusing and two-dimensional gel electrophoresis
    David B Friedman
    Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee, USA
    Methods Enzymol 463:515-40. 2009
    ..separation techniques for intact proteins in a single analytical run continues to be by the combination of isoelectric focusing (IEF) and SDS-PAGE, originally introduced by O'Farrell [(1975). J. Biol. Chem.250, 4007-4021]...
  2. pmc Peptide separations by on-line MudPIT compared to isoelectric focusing in an off-gel format: application to a membrane-enriched fraction from C2C12 mouse skeletal muscle cells
    Sarah Elschenbroich
    Ontario Cancer Institute, University Health Network, Canada
    J Proteome Res 8:4860-9. 2009
    ..Recently, Agilent Technologies has introduced the OFFGEL fractionator, which conducts peptide separation by isoelectric focusing in an off-gel setup...
  3. ncbi Laboratory testing of individuals with severe alpha1-antitrypsin deficiency in three European centres
    M Miravitlles
    Fundació Clínic, Institut d Investigacions Biomediques August Pi i Sunyer, CIBER de Enfermedades Respiratorias, Barcelona, Spain
    Eur Respir J 35:960-8. 2010
    ..determination of the level of alpha(1)-AT and genotyping are reported as the main diagnostic steps, whereas isoelectric focusing (also referred to as phenotyping) is reserved for confirmatory analysis...
  4. ncbi Kinetics of local and systemic isoforms of serum amyloid A in bovine mastitic milk
    S Jacobsen
    Department of Clinical Studies, Large Animal Surgery, Royal Veterinary and Agricultural University, Dyrlaegevej 48, DK 1870 Frederiksberg C, Copenhagen, Denmark
    Vet Immunol Immunopathol 104:21-31. 2005
    ..8, 6.2, 6.8 and 7.4 were demonstrated by denaturing isoelectric focusing. In milk three highly alkaline isoforms with apparent pI values above 9.3 appeared 12 h post-inoculation...
  5. ncbi Serum amyloid A isoforms in serum and synovial fluid from spontaneously diseased dogs with joint diseases or other conditions
    Mads Kjelgaard-Hansen
    Central Laboratory, Department of Small Animal Clinical Sciences, Faculty of Life Sciences, University of Copenhagen, Groennegaardsvej 3, DK 1870 Frederiksberg, Denmark
    Vet Immunol Immunopathol 117:296-301. 2007
    ..Expression of SAA isoforms was visualized by denaturing isoelectric focusing and Western blotting...
  6. ncbi Identification of distinct N-terminal truncated forms of prion protein in different Creutzfeldt-Jakob disease subtypes
    Gianluigi Zanusso
    Department of Neurological and Visual Sciences, Section of Neurology and Agricultural and Industrial Biotechnologies, University of Verona, 37134 Verona, Italy
    J Biol Chem 279:38936-42. 2004
    ..These data show the presence of multiple PrP(Sc) conformations in sCJD and, in addition, shed new light on the correlation between sCJD phenotypes and disease-associated PrP molecules...
  7. ncbi Current two-dimensional electrophoresis technology for proteomics
    Angelika Görg
    Department of Proteomics, Technische Universitat Munchen, Freising Weihenstephan, Germany
    Proteomics 4:3665-85. 2004
    ....
  8. pmc Animal and human multidrug-resistant, cephalosporin-resistant salmonella isolates expressing a plasmid-mediated CMY-2 AmpC beta-lactamase
    P L Winokur
    University of Iowa College of Medicine, Iowa City, Iowa, USA
    Antimicrob Agents Chemother 44:2777-83. 2000
    ..between animals and man...
  9. ncbi Quantitative membrane proteomics applying narrow range peptide isoelectric focusing for studies of small cell lung cancer resistance mechanisms
    Hanna Eriksson
    Karolinska Biomics Center, Karolinska University Hospital, Stockholm, Sweden
    Proteomics 8:3008-18. 2008
    ..One of the proteins found to be differentially expressed was Serca 2, a calcium pump located in the ER membrane that potentially could result in changes of apoptotic response toward Doxorubicin...
  10. ncbi The use of sigmoid pH gradients in free-flow isoelectric focusing of human endothelial cell proteins
    Xiao Ma
    Institute of Biomedicine, Pharmacology, Aarhus University, Aarhus, Denmark
    Electrophoresis 33:1349-55. 2012
    ..electrophoresis (FFE) provides a useful step in various prefractionation protocols, since matrix-free isoelectric focusing (FF-IEF) performed in this machine enables the enrichment of large, easily absorbable, sensitive proteins...
  11. ncbi Reduction and alkylation of proteins in preparation of two-dimensional map analysis: why, when, and how?
    B Herbert
    Proteome Systems, North Ryde, Sydney, NSW, Australia
    Electrophoresis 22:2046-57. 2001
    The standard procedure adopted up to the present in proteome analysis calls for just reduction prior to the isoelectric focusing/immobilized pH gradient (IEF/IPG) step, followed by a second reduction/alkylation step in between the first ..
  12. ncbi A comparison of immobilized pH gradient isoelectric focusing and strong-cation-exchange chromatography as a first dimension in shotgun proteomics
    Amal S Essader
    Research Triangle Institute, Research Triangle Park, NC 27709 2194, USA
    Proteomics 5:24-34. 2005
    ..This methodology employs isoelectric focusing of peptides on immobilized pH gradient (IPG) gels in the first dimension, followed by reversed-phase ..
  13. ncbi Characterization of cellobiohydrolase I (Cel7A) glycoforms from extracts of Trichoderma reesei using capillary isoelectric focusing and electrospray mass spectrometry
    J P Hu
    Institute for Biological Sciences, Ottawa, Ontario, Canada
    J Chromatogr B Biomed Sci Appl 752:349-68. 2001
    Capillary isoelectric focusing (CIEF) was used to profile the cellulase composition in complex fermentation samples of secreted proteins from Trichoderma reesei...
  14. pmc Endogenous phosphatidylcholine and a long spacer ligand stabilize the lipid-binding groove of CD1b
    Luis F Garcia-Alles
    CNRS, UMR5089, Département Mécanismes Moléculaires des Infections Mycobactériennes, Institut de Pharmacologie et de Biologie Structurale, Toulouse, France
    EMBO J 25:3684-92. 2006
    ..Using isoelectric focusing, native mass spectrometry and resolving the crystal structure at 1...
  15. pmc Outbreak of Klebsiella pneumoniae producing a new carbapenem-hydrolyzing class A beta-lactamase, KPC-3, in a New York Medical Center
    Neil Woodford
    Antibiotic Resistance Monitoring and Reference Laboratory, Specialist and Reference Microbiology Division Colindale, Health Protection Agency, London, United Kingdom
    Antimicrob Agents Chemother 48:4793-9. 2004
    ..Although producers of these enzymes remain rare, the progress of this enzyme group merits monitoring...
  16. ncbi Evaluation of sample fractionation using micro-scale liquid-phase isoelectric focusing on mass spectrometric identification and quantitation of proteins in a SILAC experiment
    Annika Thorsell
    Neurochemistry Laboratory, Institute of Neuroscience and Physiology, Goteborg University, Sahlgrenska University Hospital, Molndal, Sweden
    Rapid Commun Mass Spectrom 21:771-8. 2007
    ..We show that micro-scale liquid-phase isoelectric focusing in the Micro Rotofor separates proteins into well-defined fractions and reduces the sample complexity...
  17. pmc Evaluation of strong cation exchange versus isoelectric focusing of peptides for multidimensional liquid chromatography-tandem mass spectrometry
    Robbert J C Slebos
    Jim Ayers Institute for Precancer Detection and Diagnosis, Vanderbilt Ingram Cancer Center, Medical Research Building III, 465 21st Avenue South, Nashville, Tennessee 37232, USA
    J Proteome Res 7:5286-94. 2008
    ..by analyzing tryptic digests of whole cell and tissue proteomes using strong cation exchange (SCX) and isoelectric focusing (IEF) separations of peptides prior to LC-MS/MS analysis on a LTQ-Orbitrap hybrid instrument...
  18. ncbi Interference of transferrin isoform types with carbohydrate-deficient transferrin quantification in the identification of alcohol abuse
    A Helander
    Department of Clinical Neuroscience, Section of Clinical Alcohol and Drug Addiction Research, Karolinska Institutet, SE 171 76 Stockholm, Sweden
    Clin Chem 47:1225-33. 2001
    ..Isoforms of transferrin interfere with measurement of carbohydrate-deficient transferrin (CDT) as a marker of heavy alcohol consumption. We evaluated the rate of inaccurate CDT results by immunoassays...
  19. pmc Isoelectric focusing technology quantifies protein signaling in 25 cells
    Roger A O'Neill
    Cell Biosciences Inc, 1050 Page Mill Road, Palo Alto, CA 94304, USA
    Proc Natl Acad Sci U S A 103:16153-8. 2006
    A previously undescribed isoelectric focusing technology allows cell signaling to be quantitatively assessed in <25 cells...
  20. ncbi Two-stage Off-Gel isoelectric focusing: protein followed by peptide fractionation and application to proteome analysis of human plasma
    Manfred Heller
    DiagnoSwiss SA, CH 1870 Monthey, Switzerland
    Electrophoresis 26:1174-88. 2005
    ....
  21. ncbi A simple screening test for variant transthyretins associated with familial transthyretin amyloidosis using isoelectric focusing
    L H Connors
    Department of Medicine, Boston University School of Medicine, MA 02118, USA
    Biochim Biophys Acta 1407:185-92. 1998
    ..5% acrylamide (PAGE) followed by isoelectric focusing (IEF) between pH 4.0 and 7.0 in 2.5 M urea...
  22. ncbi On-line chemiluminescence detection for isoelectric focusing of heme proteins on microchips
    Xiangyi Huang
    College of Chemistry and Chemical Engineering, Shanghai Jiaotong University, Shanghai, P R China
    Electrophoresis 26:3595-601. 2005
    ..Cytochrome c, myoglobin, and horseradish peroxidase were well separated within 10 min on a glass chip and the detection limits (S/N=3) were 1.2x10(-7), 1.6x10(-7), and 1.0x10(-10) M, respectively...
  23. ncbi Adaptation of capillary isoelectric focusing to microchannels on a glass chip
    O Hofmann
    Vysis, Inc, Downers Grove, Illinois 60515, USA
    Anal Chem 71:678-86. 1999
    As a first step toward adaptation of capillary isoelectric focusing (cIEF) to microchannels on a glass chip, we have compared the three most common mobilization methods: chemical, hydrodynamic, and electroosmotic flow (EOF)-driven ..
  24. ncbi Unusual hepatic mitochondrial arginase in an Indian air-breathing teleost, Heteropneustes fossilis: purification and characterization
    Shilpee Srivastava
    Biochemical Adaptation Laboratory, Department of Zoology, Banaras Hindu University, Varanasi 221005, India
    Comp Biochem Physiol B Biochem Mol Biol 164:133-41. 2013
    ..fossilis hepatic ARG II showed cross reactivity with a 96 kDa band on native PAGE and a 48 kDa band on SDS-PAGE. The molecular, immunological and kinetic properties suggest uniqueness of the hepatic mitochondrial ARG II in H. fossilis...
  25. ncbi Interlaboratory reproducibility of isoelectric focusing in oligoclonal band detection
    Diego Franciotta
    Clin Chem 53:1557-8. 2007
  26. ncbi Validation of capillary zone electrophoresis and capillary isoelectric focusing separations optimized for the characterization of two recombinant products of the birch pollen allergen Bet v 1a
    Barbara Kronsteiner
    Division of Chemistry, Department of Molecular Biology, University of Salzburg, Salzburg, Austria
    Electrophoresis 29:2539-49. 2008
    ..Results demonstrate the suitability of both separation systems to discriminate between nonmodified Bet v 1a and carbamylated variants in the selected recombinant allergen products...
  27. ncbi Separation of hemoglobin variants with similar charge by capillary isoelectric focusing: value of isoelectric point for identification of common and uncommon hemoglobin variants
    J M Hempe
    Department of Pediatrics, Louisiana State University School of Medicine, New Orleans 70112, USA
    Electrophoresis 21:743-8. 2000
    ..variants with similar charge to evaluate the diagnostic sensitivity and specificity of automated capillary isoelectric focusing (CIEF). Peak separation was observed for each variant in samples containing Hb S, D, and G...
  28. ncbi Polyacrylamide gel plugs enabling 2-D microfluidic protein separations via isoelectric focusing and multiplexed sodium dodecyl sulfate gel electrophoresis
    Jikun Liu
    Department of Mechanical Engineering, University of Maryland, College Park, MD 20742, USA
    Electrophoresis 29:2241-50. 2008
    ..1%...
  29. ncbi Capillary isoelectric focusing and sodium dodecyl sulfate-capillary gel electrophoresis of recombinant humanized monoclonal antibody HER2
    G Hunt
    Department of Quality Control Clinical Development, Genentech Inc, South San Francisco, CA 94080, USA
    J Chromatogr A 744:295-301. 1996
    Capillary isoelectric focusing (cIEF) and IEF of recombinant humanized monoclonal antibody HER2 (rhuMAbHER2) show five charged isoforms with estimated pI values ranging from 8.6-9.1...
  30. ncbi Peptide separation with immobilized pI strips is an attractive alternative to in-gel protein digestion for proteome analysis
    Nina C Hubner
    Department of Proteomics and Signal Transduction, Max Planck Institute for Biochemistry, Martinsried, Germany
    Proteomics 8:4862-72. 2008
    ..Advantages of the in-gel format include better reliability and robustness. Considering its superior performance, diminished sample and work-up requirements, peptide IEF will become a method of choice for sample preparation in proteomics...
  31. ncbi Shotgun proteomics: a qualitative approach applying isoelectric focusing on immobilized pH gradient and LC-MS/MS
    Laurent Geiser
    Swiss Centre for Applied Human Toxicology, Geneva University, Geneva, Switzerland
    Methods Mol Biol 681:449-58. 2011
    ..to two orthogonal techniques: peptides are first separated according to their isoelectric point (pI) by isoelectric focusing (IEF) on immobilized pH gradient (IPG); after peptide extraction, they are then separated in the second ..
  32. ncbi Integration of isoelectric focusing with parallel sodium dodecyl sulfate gel electrophoresis for multidimensional protein separations in a plastic microfluidic [correction of microfludic] network
    Yan Li
    Department of Chemistry and Biochemistry, Institute for System Research, University of Maryland, College Park, MD 20742, USA
    Anal Chem 76:742-8. 2004
    An integrated protein concentration/separation system, combining non-native isoelectric focusing (IEF) with sodium dodecyl sulfate (SDS) gel electrophoresis on a polymer microfluidic chip, is reported...
  33. ncbi Paraneoplastic antibodies detected by isoelectric focusing of cerebrospinal fluid and serum
    Anette Storstein
    Department of Neurology, Haukeland University Hospital, N 5021, Bergen, Norway
    J Neuroimmunol 155:150-4. 2004
    Patients with paraneoplastic neurological syndromes often produce intrathecal antibodies. We have employed isoelectric focusing and peroxidase-labeled anti-IgG or 35S-labeled Hu or Yo antigens to identify oligoclonal bands (OCB) ..
  34. ncbi Proteolysis during the isoelectric focusing step of two-dimensional gel electrophoresis may be a common problem
    Christine Finnie
    Department of Chemistry, Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK 2500 Valby, Copenhagen, Denmark
    Anal Biochem 311:182-6. 2002
  35. pmc A proteomics platform combining depletion, multi-lectin affinity chromatography (M-LAC), and isoelectric focusing to study the breast cancer proteome
    Zhi Zeng
    Department of Chemistry and Chemical Biology, Barnett Institute, Northeastern University, Boston, Massachusetts, USA
    Anal Chem 83:4845-54. 2011
    ..The collected glycoproteomes were then subjected to isoelectric focusing (IEF) separation by a digital ProteomeChip (dPC), followed by in-gel digestion and LC-MS analysis using an ..
  36. ncbi Identification of proteins in human cerebrospinal fluid using liquid-phase isoelectric focusing as a prefractionation step followed by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionisation mass spectrometry
    P Davidsson
    Department of Clinical Neuroscience, Experimental Neuroscience Section, Goteborg University, Sahlgrenska University Hospital Molndal, Sweden
    Rapid Commun Mass Spectrom 16:2083-8. 2002
    ..We have developed a method in which liquid-phase isoelectric focusing (IEF) is used to prefractionate individual CSF samples; selected IEF fractions are then analysed on SYPRO-..
  37. ncbi Integrated platform of capillary isoelectric focusing, trypsin immobilized enzyme microreactor and nanoreversed-phase liquid chromatography with mass spectrometry for online protein profiling
    Tingting Wang
    Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, PP R China
    Electrophoresis 32:2848-56. 2011
    ..We anticipate that the integrated platform developed herein will provide a promising tool for low-abundance protein identification with the combination of top-down and bottom-up approaches...
  38. ncbi Charge heterogeneity of monoclonal antibodies by multiplexed imaged capillary isoelectric focusing immunoassay with chemiluminescence detection
    David A Michels
    Department of Protein Analytical Chemistry, Genentech, A Member of the Roche Group, 1 DNA Way, South San Francisco, California 94080, USA
    Anal Chem 84:5380-6. 2012
    ..This assay offers specificity, speed and sensitivity advantages over conventional capillary isoelectric focusing (CIEF) platforms...
  39. ncbi Capillary electrophoresis and isoelectric focusing in peptide and protein analysis
    Pier Giorgio Righetti
    Miles Gloriosus Academy, Milano, Italy
    Proteomics 13:325-40. 2013
    ..To this purpose, commercial instrumentation is already available on the market. This tutorial is part of the International Proteomics Tutorial Programme (IPTP 13)...
  40. pmc Two-dimensional capillary electrophoresis: capillary isoelectric focusing and capillary zone electrophoresis with laser-induced fluorescence detection
    Jane A Dickerson
    Department of Chemistry, University of Washington, Seattle WA, USA
    Electrophoresis 31:2650-4. 2010
    ..A fraction was transferred to the second-dimension capillary for separation. The process of fraction transfer and second dimension separation was repeated two dozen times. The separation produced a spot capacity of 125...
  41. ncbi Continuous mode of operation for large volume dosing in analytical carrier ampholyte-free isoelectric focusing of proteins applied to off-line detection of fractions
    Jana Budilová
    Masaryk University, Department of Analytical Chemistry, Brno, Czech Republic
    J Sep Sci 29:1613-21. 2006
    ..A useful method for this purpose is Carrier Ampholyte Free-Isoelectric Focusing (CAF-IEF)...
  42. ncbi Characterization of the phosphoproteome in LNCaP prostate cancer cells by in-gel isoelectric focusing and tandem mass spectrometry
    Li Chen
    Department of Pharmaceutical Sciences, Charles B Stout Neuroscience Mass Spectrometry Laboratory, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA
    J Proteome Res 9:174-8. 2010
    ..The analytical methodology used in the study included separation of the LNCaP proteins by in-gel isoelectric focusing (IEF), digestion of the proteins with trypsin, enrichment of the digests for phosphopeptides with ..
  43. ncbi Immunological and proteomic analysis of preparative isoelectric focusing separated culture filtrate antigens of Mycobacterium tuberculosis
    Anbarasu Deenadayalan
    Department of Immunology, Tuberculosis Research Centre ICMR, Mayor V R Ramanathan Road, Chetput, Chennai 600 031, India
    Exp Mol Pathol 88:156-62. 2010
    ..tuberculosis. In this study, preparative liquid-phase isoelectric focusing was used for the separation of culture filtrate protein (CFP) of M. tuberculosis...
  44. ncbi Fast isoelectric focusing and antipeptide antibodies for detecting bovine casein in adulterated water buffalo milk and derived mozzarella cheese
    Francesco Addeo
    Dipartimento di Scienza degli Alimenti, Universita degli Studi di Napoli Federico II, Via Universita 100, Parco Gussone, I 80055 Portici, Italy
    J Agric Food Chem 57:10063-6. 2009
    ..After isoelectric focusing of CN plasminolysates, performed according to the official method, immunoblot analysis with antipeptide ..
  45. ncbi A possible role of intracellular isoelectric focusing in the evolution of eukaryotic cells and multicellular organisms
    Jaroslav Flegr
    Department of Philosophy and History of Science, Charles University, Vinicna 7, 128 44, Praha 2, Czech Republic
    J Mol Evol 69:444-51. 2009
    ..5 kV/cm, the value sufficient for effective isoelectric focusing of cytoplasmic proteins...
  46. ncbi Determination of the isoelectric point of proteins by capillary isoelectric focusing
    Pier Giorgio Righetti
    Department of Agricultural and Industrial Biotechnology, University of Verona, Strada Le Grazie 15, Verona 37134, Italy
    J Chromatogr A 1037:491-9. 2004
    Different ways of determining isoelectric points (pI) of proteins in capillary isoelectric focusing are reviewed here...
  47. ncbi Screening for congenital disorders of glycosylation (CDG): transferrin HPLC versus isoelectric focusing (IEF)
    Ester Quintana
    Institut de Bioquimica Clinica, Servei de Bioquímica i Genètica Molecular, Hospital Clinic, Barcelona, Spain
    Clin Biochem 42:408-15. 2009
    ..We present our experience and advantages of the implementation of Tf-HPLC...
  48. pmc Electrolytic reduction: modification of proteins occurring in isoelectric focusing electrophoresis and in electrolytic reactions in the presence of high salts
    Der Yen Lee
    Graduate Institute of Biochemical Sciences, College of Life Science, National Taiwan University, P O Box 23 106, Taipei 106, Taiwan
    Anal Chem 81:3957-64. 2009
    Artifacts in two-dimensional electrophoresis (2-DE) caused by the presence of salts in isoelectric focusing (IEF) have been previously described as a result of increasing conductivity and inducing electroosmosis...
  49. ncbi Phosphoproteome analysis by in-gel isoelectric focusing and tandem mass spectrometry
    Sarka Beranova-Giorgianni
    Department of Pharmaceutical Sciences, The University of Tennessee Health Science Center, 5P Crowe Bldg, 874 Union Avenue, Memphis, TN 38163, USA
    Methods Mol Biol 519:383-96. 2009
    ..This chapter describes an in-gel isoelectric focusing-liquid chromatography-tandem mass spectrometry (IEF-LC-MS/MS) analytical strategy for phosphoproteome ..
  50. ncbi Evaluation of protein loading techniques and improved separation in OFFGEL isoelectric focusing
    Eva Maria Keidel
    Max Planck Institute of Biochemistry, Protein Analysis Group, Martinsried, Germany
    Electrophoresis 32:1659-66. 2011
    ..Second, a 48-well OFFGEL frame was developed, which doubles the resolution of the commercially available 24-well frame. It is capable of separating proteins with small pI differences and shows potential for isoform/PTM separation...
  51. ncbi Oligoclonal IgG bands in Japanese patients with multiple sclerosis. A comparative study between isoelectric focusing with IgG immunofixation and high-resolution agarose gel electrophoresis
    Ichiro Nakashima
    Department of Neurology, Tohoku University School of Medicine, 1 1 Seiryo machi, Aoba ku, Sendai 980 8574, Japan
    J Neuroimmunol 159:133-6. 2005
    ..We compared the frequency of OB between isoelectric focusing (IEF) and agarose gel electrophoresis (AGE) in 59 Japanese patients with clinically definite MS [39 with ..
  52. ncbi Genetic analysis of Bemisia (Hemiptera: Aleyrodidae) populations by isoelectric focusing electrophoresis
    J K Brown
    Department of Plant Sciences, University of Arizona, Tucson 85721, USA
    Biochem Genet 38:13-25. 2000
    ..The latter two groups were grouped separately from New World populations and are thought to have a recent origin in the Eastern Hemisphere...
  53. ncbi Isoelectric focusing as a tool for the investigation of post-translational processing and chemical modifications of proteins
    E Gianazza
    Istituto di Scienze Farmacologiche, Milan, Italy
    J Chromatogr A 705:67-87. 1995
    ..As an important field of application of the above concepts may be genetic engineering, an exhaustive bibliographic list dealing with pI evaluation and structural assessment on recombinant proteins is included...
  54. pmc Isoelectric focusing in a drop
    Noah G Weiss
    Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287 9709, United States
    Langmuir 27:494-8. 2011
    A novel approach to molecular separations is investigated using a technique termed droplet-based isoelectric focusing. Drops are manipulated discretely on a superhydrophobic surface, subjected to low voltages for isoelectric focusing, and ..
  55. ncbi A simple and rapid technique for detecting protein phosphorylation using one-dimensional isoelectric focusing gels and immunoblot analysis
    Jeffrey C Anderson
    Department of Biochemistry, University of Missouri Columbia, Columbia, MO, USA
    Plant J 55:881-5. 2008
    We report a technique for detecting protein phosphorylation that involves isoelectric focusing in a vertical mini-gel format followed by immunoblot detection of the target protein...
  56. ncbi A reference map of human lung MRC-5 fibroblast proteins using immobilized pH gradient-isoelectric focusing-based two-dimensional electrophoresis
    K Y Leung
    National Heart and Lung Institute, Imperial College School of Medicine, Heart Science Centre, Harefield Hospital, Harefield, Middlesex, UB9 6JH, UK
    Proteomics 1:787-94. 2001
    We report the first protein map of human adult lung MRC-5 fibroblasts using isoelectric focusing-immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis...
  57. pmc Examining serum amyloid P component microheterogeneity using capillary isoelectric focusing and MALDI-MS
    Noah G Weiss
    Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287 1604, USA
    Proteomics 11:106-13. 2011
    ..Overall, this work corroborates the idea that SAP is relatively consistent across the population and with respect to microheterogeneity...
  58. ncbi Characterisation of stage-specific proteins of Oesophagostomum dentatum by preparative isoelectric focusing and lectin blotting
    A Joachim
    Institut für Parasitologie, Tierarztliche Hochschule Hannover, Bunteweg 17, D 30559, Hannover, Germany
    Parasitol Int 50:41-5. 2001
    ..to their protein and glycoprotein patterns by solubility-based protein fractionation and preparative isoelectric focusing followed by SDS-PAGE or by Western blotting with various lectins...
  59. ncbi Capillary isoelectric focusing of the scrapie prion protein
    M J Schmerr
    US Department of Agriculture, National Animal Disease Center, Ames, IA 50010, USA
    J Chromatogr A 802:135-41. 1998
    ..In this study, we explored the use of capillary isoelectric focusing (cIEF) to characterize the pI values for PrPsc isolated from sheep and hamster brain...
  60. ncbi Free-flow isoelectric focusing of proteins remaining in cell fragments following sonication of thyroid carcinoma cells
    Christian Obermaier
    FFE Weber GmbH, Martinsried, Germany
    Electrophoresis 26:2109-16. 2005
    ..even those proteins which remained in cell fragments, could completely be dissolved by free-flow isoelectric focusing media. They migrated through the free-flow electrophoresis chamber without forming precipitates...
  61. ncbi Structure-based engineering of a monoclonal antibody for improved solubility
    Sheng Jiun Wu
    Biologics Research, Centocor R and D, 145 King of Prussia Radnor, PA 19087 4557, USA
    Protein Eng Des Sel 23:643-51. 2010
    ..All three engineering approaches led to improved solubility and adding an N-linked carbohydrate to the CDR was the most effective route for enhancing the solubility of CNTO607...
  62. ncbi Rapid protein separations in ultra-short microchannels: microchip sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing
    Jongyoon Han
    Sandia National Laboratories, Livermore, CA, USA
    J Chromatogr A 1049:205-9. 2004
    ..gel electrophoresis techniques, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing (IEF) in short microchannels (approximately millimeters) that take less than a minute...
  63. ncbi Fractionation of complex protein mixtures by liquid-phase isoelectric focusing
    Julie Hey
    Bio Rad Laboratories, Inc, Hercules, California, USA
    Methods Mol Biol 424:225-39. 2008
    ..The MicroRotofor isoelectric focusing (IEF) cell fractionates proteins in free solution according to their isoelectric point (pI)...
  64. ncbi Isoelectric focusing in long immobilized pH gradient gels to improve protein separation in proteomic analysis
    Julia Poland
    Institut für Laboratoriums medizin und Pathobiochemie, Universitatsklinikum Charite, Medizinische Fakultät der Humbold Universität zu Berlin, Germany
    Electrophoresis 24:1271-5. 2003
    ..We have developed an extremely long isoelectric focusing (IEF) system aimed at more complete protein profiling...
  65. ncbi Carbohydrate analysis of transferrin subfractions isolated by preparative isoelectric focusing in immobilized pH gradients
    G de Jong
    Department of Chemical Pathology, Erasmus University Rotterdam, The Netherlands
    Electrophoresis 13:225-8. 1992
    Analytical isoelectric focusing of human serum transferrin has revealed the presence of up to nine different transferrin subfractions...
  66. ncbi Clinical and diagnostic approach in unsolved CDG patients with a type 2 transferrin pattern
    M Mohamed
    Institute for Genetic and Metabolic Diseases at the Department of Pediatrics, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
    Biochim Biophys Acta 1812:691-8. 2011
    ....
  67. ncbi Separation of human cerebrospinal fluid proteins by capillary isoelectric focusing in the absence of denaturing agents
    T Manabe
    Department of Chemistry, Faculty of Science, Ehime University, Matsuyama, Japan
    Electrophoresis 20:3677-83. 1999
    Conditions of capillary isoelectric focusing (CIEF) to separate human cerebrospinal fluid (CSF) proteins were examined referring to those which we have established for the separation of human plasma/serum proteins...
  68. ncbi Isoelectric focusing in immobilized pH gradients: recent analytical and preparative developments
    P G Righetti
    Department of Agricultural and Industrial Biotechnologies, University of Verona, Italy
    Anal Biochem 247:1-10. 1997
    b>Isoelectric focusing in immobilized pH gradients (IPG), covering both analytical and preparative aspects, is here reviewed...
  69. ncbi High-efficiency capillary isoelectric focusing of protein complexes from Escherichia coli cytosolic extracts
    Y Shen
    Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA
    J Chromatogr A 914:257-64. 2001
    High-efficiency capillary isoelectric focusing (cIEF) separations of protein complexes obtained from soluble protein fractions are demonstrated...
  70. ncbi Proteome analysis of microdissected tumor tissue using a capillary isoelectric focusing-based multidimensional separation platform coupled with ESI-tandem MS
    Yueju Wang
    Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA
    Anal Chem 77:6549-56. 2005
    ..Capillary isoelectric focusing combined with nano-reversed-phase liquid chromatography in an automated and integrated platform not only ..
  71. ncbi An optimized procedure for detection of proteins on carrier ampholyte isoelectric focusing and immobilized pH gradient gels with imidazole and zinc salts: its application to the identification of isoelectric focusing separated isoforms by in-gel proteolys
    L Castellanos-Serra
    Division of Physical Chemistry, Center for Genetic Engineering and Biotechnology, Havana, Cuba
    Electrophoresis 22:1677-85. 2001
    A method for the characterization of proteins separated by isoelectric focusing in carrier ampholytes (CA-IEF) or immobilized pH gradient (IPG) gels by in-gel digestion and mass spectrometry is described...
  72. ncbi Transferrin D protein variants in the diagnosis of congenital disorders of glycosylation (CDG)
    Eliska Marklová
    Department of Pediatrics, University Hospital, Sokolska 581, Hradec Kralove, Czech Republic
    J Clin Lab Anal 23:77-81. 2009
    ..Analysis of serum Tf by isoelectric focusing is used as a common method suitable for screening 19 out of a total of 22 types of glycosylation defects ..
  73. ncbi Development of a multilane channel system for nongel-based two-dimensional protein separations using isoelectric focusing and asymmetrical flow field-flow fractionation
    Ki Hun Kim
    Department of Chemistry, Yonsei University, Seoul, 120 749, Korea
    Anal Chem 81:1715-21. 2009
    A dual purpose multilane channel system to carry out isoelectric focusing (IEF) and asymmetrical flow field-flow fractionation (IEF-AFlFFF or IEF-AF4) was developed for the high-speed fractionation of a proteome in two dimensions (2D): ..
  74. ncbi High speed two-dimensional protein separation without gel by isoelectric focusing-asymmetrical flow field flow fractionation: application to urinary proteome
    Ki Hun Kim
    Department of Chemistry, Yonsei University, Seoul, Korea
    J Proteome Res 8:4272-8. 2009
    An online multilane channel system for isoelectric focusing and asymmetrical flow field-flow fractionation (IEF-AF4) is utilized for the two-dimensional separation (2D: isoelectric point, pI, and hydrodynamic diameter, d(s)) of a human ..
  75. ncbi Characterization of the rat liver membrane proteome using peptide immobilized pH gradient isoelectric focusing
    J M Chick
    Department of Chemistry and Biomolecular Sciences, Macquarie University, NSW, 2109, Australia
    J Proteome Res 7:1036-45. 2008
    ..prior to analysis using a gel-based shotgun proteomics approach called peptide immobilized pH gradient isoelectric focusing (IPG-IEF)...
  76. ncbi Development of non-gel-based two-dimensional separation of intact proteins by an on-line hyphenation of capillary isoelectric focusing and hollow fiber flow field-flow fractionation
    Dukjin Kang
    Department of Chemistry, Yonsei University, Seoul, 120 749, Korea
    Anal Chem 78:5789-98. 2006
    ..by first exploiting the differences in their respective isoelectric points (pI) in a Teflon capillary using isoelectric focusing (IEF), followed by a molecular weight (MW)-based separation in a hollow fiber by flow field-flow ..
  77. ncbi Computer-assisted evaluation of isoelectric focusing patterns in electrophoretic gels: identification of smoothhounds (Mustelus mustelus, Mustelus asterias) and comparison with lower value shark species
    P Renon
    Istituto di Ispezione degli Alimenti di Origine Animale, , , Italy
    Electrophoresis 22:1534-8. 2001
    ..Mustelus mustelus) and starry smoothhound (Mustelus asterias), have been identified by polyacrylamide isoelectric focusing (IEF-PAGE), along with several shark species of minor commercial value...
  78. ncbi Recycling isoelectric focusing and isotachophoresis
    M Bier
    University of Arizona, Tucson 85721, USA
    Electrophoresis 19:1057-63. 1998
    Of all electrophoretic methods, isoelectric focusing offers the highest resolution and is best suited for preparative applications...
  79. ncbi Capillary isoelectric focusing-based multidimensional concentration/separation platform for proteome analysis
    Jinzhi Chen
    Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA
    Anal Chem 75:3145-52. 2003
    An integrated proteome concentration/separation approach involving on-line combination of capillary isoelectric focusing (CIEF) with capillary reversed-phase liquid chromatography (CRPLC) is developed for providing significant analyte ..
  80. ncbi Gel based isoelectric focusing of peptides and the utility of isoelectric point in protein identification
    Benjamin J Cargile
    Mass Spectrometry Program, Research Triangle Institute, 3040 Cornwallis Road, Research Triangle Park, North Carolina 27709, USA
    J Proteome Res 3:112-9. 2004
    ..This approach was successfully applied to analysis of the soluble fraction of the E. coli proteome, where 417 proteins were identified from 1022 peptides using just 20 microg of material...
  81. ncbi iTRAQ compatibility of peptide immobilized pH gradient isoelectric focusing
    Johan Lengqvist
    Karolinska Biomic Center, Karolinska University Hospital, Stockholm, Sweden
    Proteomics 7:1746-52. 2007
    Immobilized pH gradient isoelectric focusing (IPG-IEF) has emerged as a highly promising alternative to strong-cation exchange fractionation as the first dimension in shot-gun proteomics...
  82. ncbi Large scale protein profiling by combination of protein fractionation and multidimensional protein identification technology (MudPIT)
    Emily I Chen
    Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA
    Mol Cell Proteomics 5:53-6. 2006
    ..As a result, we were able to reduce sample complexity by protein fractionation and increase the possibility of detecting proteins with lower abundance in the complex protein mixture...
  83. ncbi Congenital disorders of glycosylation: review of their molecular bases, clinical presentations and specific therapies
    T Marquardt
    Klinik und Poliklinik für Kinderheilkunde, Albert Schweitzer Str 33, 48149 Munster, Germany
    Eur J Pediatr 162:359-79. 2003
    ..Since the clinical spectrum of symptoms in CDG is variable and may be unspecific, a generous selective screening for the presence of CDG is recommended...
  84. ncbi Isoelectric focusing in serial immobilized pH gradient gels to improve protein separation in proteomic analysis
    Slobodan Poznanovic
    ProteoSys AG, Carl Zeiss Strasse 51, D 55129 Mainz, Germany
    Electrophoresis 26:3185-90. 2005
    We previously demonstrated the separation of proteins by isoelectric focusing (IEF) over pH 4-8 immobilized pH gradients (IPGs) over 54 cm (Poland et al., Electrophoresis 2003, 24, 1271)...
  85. ncbi Identification of synaptic vesicle, pre- and postsynaptic proteins in human cerebrospinal fluid using liquid-phase isoelectric focusing
    P Davidsson
    Department of Clinical Neuroscience, Sahlgrenska University Hospital Molndal, Sweden
    Electrophoresis 20:431-7. 1999
    ..a procedure including precipitation of CSF proteins with trichloroacetic acid, followed by liquid-phase isoelectric focusing using the Rotofor Cell, and finally analysis of Rotofor fractions by Western blotting for identification of ..
  86. ncbi Matching of oligoclonal immunoglobulin transcriptomes and proteomes of cerebrospinal fluid in multiple sclerosis
    Birgit Obermeier
    Institute of Clinical Neuroimmunology, Ludwig Maximilians University, University Hospital Grosshadern, Marchioninistrasse 15, D 81377 Munich, Germany
    Nat Med 14:688-93. 2008
    ..This approach can be applied to other organ-specific diagnostic fluid or tissue samples to compare the Ig transcripts of local B cells with the corresponding antibody proteomes of individual subjects...
  87. pmc Immobilized pH gradient isoelectric focusing as a first-dimension separation in shotgun proteomics
    Benjamin J Cargile
    Research Triangle Institute, 3040 Cornwallis Road, Research Triangle Park, NC 27709 2194, USA
    J Biomol Tech 16:181-9. 2005
    ..of an alternative first-dimension separation technique for shotgun proteomics, immobilized pH gradient isoelectric focusing (IPG-IEF)...
  88. ncbi Stabilization of hypoglycosylation in a patient with congenital disorder of glycosylation type Ia
    S H Hahn
    Department of Laboratory Medicine, Mayo Clinic College of Medicine, Rochester, Minnesota, USA
    J Inherit Metab Dis 29:235-7. 2006
    ..b>Isoelectric focusing for serum transferrin is a widely used screening method but there could be a limit of detection and the ..
  89. ncbi Proteome analysis of human plasma and amniotic fluid by Off-Gel isoelectric focusing followed by nano-LC-MS/MS
    Philippe E Michel
    DiagnoSwiss SA, Monthey, Switzerland
    Electrophoresis 27:1169-81. 2006
    ..This representation shows that 36 of 49 reference proteins could be identified in both data sets, and that isoform shifts in pI are well conserved in the OGE data sets...
  90. ncbi Diagnosis of congenital disorders of glycosylation by capillary zone electrophoresis of serum transferrin
    Hubert A Carchon
    Department of Pediatrics, Center for Metabolic Disease, Katholieke Universiteit Leuven, B 3000 Leuven, Belgium
    Clin Chem 50:101-11. 2004
    Congenital disorders of glycosylation (CDG) are usually diagnosed by isoelectric focusing (IEF) of serum transferrin (Tf). The aim of this study was to evaluate capillary zone electrophoresis (CZE) as a diagnostic alternative to IEF.
  91. ncbi Evaluating preparative isoelectric focusing of complex peptide mixtures for tandem mass spectrometry-based proteomics: a case study in profiling chromatin-enriched subcellular fractions in Saccharomyces cerevisiae
    Hongwei Xie
    Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 321 Church Street SE, 6 155 Jackson Hall, Minneapolis, Minnesota 55455, USA
    Anal Chem 77:3198-207. 2005
    We have evaluated the use of free-flow electrophoresis, an emerging separation method for preparative isoelectric focusing of complex peptide mixtures, as a tool for high-throughput tandem mass spectrometry-based proteomic analysis...
  92. ncbi Comprehensive analysis of complex proteomes using microscale solution isoelectrofocusing prior to narrow pH range two-dimensional electrophoresis
    Xun Zuo
    The Wistar Institute, Philadelphia, PA 19104, USA
    Proteomics 2:58-68. 2002
    ..of this strategy is to prefractionate samples into a few well-defined pools using microscale solution isoelectric focusing (nusol-IEF) prior to 2-DE protein analysis...
  93. ncbi Added value for tandem mass spectrometry shotgun proteomics data validation through isoelectric focusing of peptides
    Manfred Heller
    DiagnoSwiss SA, Monthey, Switzerland
    J Proteome Res 4:2273-82. 2005
    ..We replaced the first dimension by isoelectric focusing in the liquid phase using the Off-Gel device producing 15 fractions...
  94. ncbi Efficient separation and analysis of peroxisomal membrane proteins using free-flow isoelectric focusing
    Gerhard Weber
    TECAN Munich, Proteomics Division, Kirchheim, Germany
    Electrophoresis 25:1735-47. 2004
    ..hydrophobic proteins in solution, an urea/thiourea/detergent mixture, as used in traditional gel-based isoelectric focusing (IEF), was added to the electrophoresis buffer...
  95. ncbi Preparative peptide isoelectric focusing as a tool for improving the identification of lysine-acetylated peptides from complex mixtures
    Hongwei Xie
    Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455, USA
    J Proteome Res 6:2019-26. 2007
    ..In a proof-of-principle study of whole cell yeast lysates, we demonstrate the combination of preparative isoelectric focusing using free-flow electrophoresis, and an adjusted peptide isoelectric point prediction algorithm, as an ..
  96. ncbi Comparison of the isoelectric focusing patterns of darbepoetin alfa, recombinant human erythropoietin, and endogenous erythropoietin from human urine
    Don H Catlin
    UCLA Olympic Analytical Laboratory, Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90025, USA
    Clin Chem 48:2057-9. 2002
  97. ncbi Rapid determination of transferrin isoforms by immunoaffinity liquid chromatography and electrospray mass spectrometry
    J M Lacey
    Biochemical Genetics Laboratory, Department of Laboratory Medicine and Pathology, Biomedical Mass Spectrometry and Functional Proteomics Facility, Mayo Clinic, 200 First St. SW, Rochester, MN 55905, USA
    Clin Chem 47:513-8. 2001
    ..Methods to resolve CDT from fully glycosylated transferrin (Trf) have been based on a neutral shift in the isoelectric focusing (IEF) pattern or on a reduction in the negative charge, allowing resolution by anion-exchange ..
  98. ncbi Combining microscale solution-phase isoelectric focusing with Multiplexed Proteomics dye staining to analyze protein post-translational modifications
    Birte Schulenberg
    Proteomics Department, Molecular Probes, Inc, Eugene, OR 97402, USA
    Electrophoresis 25:2539-44. 2004
    ..We determined that prefractionating samples by solution-phase isoelectric focusing is an effective alternative to sucrose-gradient fractionation that can be applied equally well to the ..
  99. ncbi Low-molecular-weight human serum proteome using ultrafiltration, isoelectric focusing, and mass spectrometry
    Robert G Harper
    Protea Biosciences Inc, West Virginia University, Department of Chemistry, Morgantown, USA
    Electrophoresis 25:1299-306. 2004
    ..complex nature of the sample was achieved by fractionation of the LMW serum proteins using solution-phase isoelectric focusing (IEF)...
  100. pmc Proteomic analysis of age dependent nitration of rat cardiac proteins by solution isoelectric focusing coupled to nanoHPLC tandem mass spectrometry
    Sung Jung Hong
    Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS 66047, USA
    Exp Gerontol 42:639-51. 2007
    ..Am. J. Physiol. Heart Circ. Physiol. 288, H371-381]. In the present report, we used solution phase isoelectric focusing (IEF) followed by nanoHPLC-ESI-MS/MS that allowed us to obtain good MS/MS data to identify specific sites ..
  101. ncbi Serum biomarker discovery in renal cancer using 2-DE and prefractionation by immunodepletion and isoelectric focusing; increasing coverage or more of the same?
    Naveen S Vasudev
    Cancer Research UK Clinical Centre, Leeds Institute of Molecular Medicine, St James s University Hospital, Leeds, UK
    Proteomics 8:5074-85. 2008
    ....

Research Grants63

  1. Chymase-Mediated MMP Activation in Ishemia Reperfusion Injury
    LOUIS JOSEPH DELL'ITALIA; Fiscal Year: 2013
    ..Nuclear protein extraction from I/R cardiomyocytes will be separated by isoelectric focusing (IEF). After IEF, proteins on agarose gels will be separated using 2D zymography...
  2. MECHANISMS OF LIVER INJURY BY HEPATITIS C AND ALCOHOL
    Steven A Weinman; Fiscal Year: 2013
    ..Methods to be employed include a novel isoelectric focusing assay for FOXO3 PTMs, examination of liver explants samples from patients undergoing liver transplantation ..
  3. Novel High Affinity and Long-Acting Bovine FSH Analogs for Veterinary Superovulat
    BRUCE DALE WEINTRAUB; Fiscal Year: 2012
    ..previously validated bFSH analog ELISA immunoassay, robust in vitro bioassay, SDS-PAGE electrophoresis and isoelectric focusing gel analysis. 3...
  4. Using PET isotopes to identify drug targets and to diagnose infectious diseases
    Peter J Tonge; Fiscal Year: 2010
    ..and fractionated under non-denaturing conditions using methods such as size-exclusion chromatography and isoelectric focusing with in-line radiation detection to identify fractions containing protein-drug complexes...
  5. Regulation of mitochondrial metabolism by lysine acetylation
    Eric S Goetzman; Fiscal Year: 2013
    ..Specific Aim 3b will use preparative isoelectric focusing to separate differentially acetylated VLCAD, MCAD, IVD and ETF isoforms from mouse liver...
  6. Phase 2 Study of rAAV1-CB-hAAT for Treatment of Alpha-1 Antitrypsin Deficiency
    Jeffrey D Chulay; Fiscal Year: 2013
    ..by evaluation of serum concentrations of M-specific AAT and total AAT and serum AAT phenotype determined on isoelectric focusing gels...
  7. cGMP manufacturing of PBI-220, a broad spectrum immunoadhesin therapeutic against
    LLOYD M YU; Fiscal Year: 2013
    ..identity, strength, potency, purity and stability will be evaluated using SDS-PAGE and immunoblotting, isoelectric focusing, UV spectroscopy, binding assays, size exclusion and peptide mapping liquid chromatography...
  8. Fast, High-Resolution Protein Separations using Controllable Electro-Osmotic Flow
    FRANK JAHNKE; Fiscal Year: 2010
    ..The proposed microfluidic device combines capillary isoelectric focusing (cIEF) with a single fast capillary zone electrophoresis (CZE)...
  9. Eukaryotic Post-Translational Modifications
    Neil L Kelleher; Fiscal Year: 2013
    ..We now propose a combination of solution isoelectric focusing (sIEF) and gel eluted liquid fraction entrapment electrophoresis (GELFrEE) prior to nanocapillary LC-MS/MS ..
  10. Two-Dimensional Electrophoresis Device for Biomarker Detection
    Z Hugh Fan; Fiscal Year: 2010
    ..limitations, we will develop a 2D electrophoresis device consisting of one channel for the first dimension (isoelectric focusing) and a number of orthogonal channels for the second dimension (polyacrylamide gel electrophoresis)...
  11. New Materials for High Efficiency Protein Separations
    Mary J Wirth; Fiscal Year: 2011
    ..We propose to achieve on-chip integration of 2D separations: isoelectric focusing followed by sieving separation in the same colloidal crystal...
  12. Mass Spectrometry Imaging of Protein Gels
    Rachel R Ogorzalek Loo; Fiscal Year: 2013
    ..Intact proteins separated by isoelectric focusing gel electrophoresis will be imaged from immobilized pH gradient (IPG) polyacrylamide gels (the first ..
  13. MECHANISMS & INHIBITORS OF PYRIDOXAL-DEPENDENT ENZYMES
    DAVID METZLER; Fiscal Year: 1993
    ..3) We will separate individual species of the self-inactivated beta subform of AspAT by isoelectric focusing and will crystallize at least the major species and some other less abundant forms...
  14. ACTIVE SPECIFIC IMMUNOTHERAPY IN MAN--A MURINE MODEL
    Barry Kahan; Fiscal Year: 1992
    ..During the previous grant period TSTA, extracted with 1-butanol and partially purified by preparative isoelectric focusing, was demonstrated to potentiate the therapeutic effects of a variety of immunostimulants: (1) ..
  15. REGULATION OF ORNITHINE DECARBOXYLASE ACTIVITY
    John Mitchell; Fiscal Year: 1980
    ..sulfate precipitation, ion exchange and molecular sieve column chromatography, SDS gel electrophoresis, and isoelectric focusing)...
  16. IMMUNOGENETICS OF MARROW ALLOGRAFTING
    John Hansen; Fiscal Year: 1992
    ..The research question addressed in Project 6 is whether the microvariants defined by individual HLA alleles have a significant impact on GVHD...
  17. BOVINE LEUKEMIA VIRUS
    DON GRAVES; Fiscal Year: 1980
    ..gel filtration in the presence of guanidine-HCL, affinity column and ion exchange chromatography, and/or isoelectric focusing. Monospecific sera will be used in the antigenic analysis of the BLV proteins as well as to determine their ..
  18. CHARACTERIZATION OF SERUM FACTORS INDUCING MITOSIS
    HARRY ANTONIADES; Fiscal Year: 1980
    ..treatment of washed platelets, followed by ion-exchange chromatography, gel filtration in l M acetic acid, isoelectric focusing and preparative SDS polyacrylamide gel electrophoresis. PDGF has a PI of about 9...
  19. PROTEASE INHIBITORS IN HUMAN BLADDER CANCER INVASION
    Brian Liu; Fiscal Year: 1993
    ..the activity of plasma membrane bound cathepsin B at a biochemical level using enzyme overlay membranes and isoelectric focusing of subcellular fractions...
  20. cIEF of glycoproteins in short nanoporous channels
    Mary J Wirth; Fiscal Year: 2013
    ..We propose to determine the feasibility of 3-mm channels packed with nonporous silica for rapid isoelectric focusing of the glycoforms of an immunoextracted protein from serum...
  21. DEVELOPMENT OF SMALL INTESTINAL EPITHELIUM
    Kwo Yih Yeh; Fiscal Year: 1990
    ..by the techniques of immunofluorescent staining, immunohistochemistry, rocket immunoelectrophoresis, isoelectric focusing, immunoblotting and in vitro mRNA translation...
  22. ASPERGILLUS FUMIGATUS COMPLEMENT INHIBITOR
    RONALD WASHBURN; Fiscal Year: 1991
    ..be chromatographically purified, and homogeneity of the final material will be judged using SDS-PAGE and isoelectric focusing. Following purification, aminoacid and monosaccharide content will be determined...
  23. GENETICS OF ANTIBODIES TO PERIODONTAL PATHOGENS
    JEROME HABER; Fiscal Year: 1990
    ..Experiments will be done in inbred mice and isoelectric focusing and idiotype specific ELISA's will be used to determine the spectrotypes and idiotypes of anti-T1 ..
  24. MECHANISMS OF RENAL URATE TRANSPORT
    Ruth Abramson; Fiscal Year: 1990
    ..gel filtration, sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE), gradient PAGE, and isoelectric focusing. When purified, polyclonal antibodies will be raised and the effect of the antibodies on urate transport in ..
  25. DERMATITIS HERPETIFORMIS & THE MUCOSAL IMMUNE RESPONSE
    Russell Hall; Fiscal Year: 1992
    ..The relationship between serum and secretory antibodies to dietary antigens will be determined by comparing isoelectric focusing (IEF) spectrotypes and analysis of idiotypic (Id) specificity...
  26. SERUM PROTEIN BINDING THE HN AUTOANTIGEN
    JOHN KANALAS; Fiscal Year: 1993
    ..Analysis of the samples will be by ELISA and SDS-PAGE/Western analysis and isoelectric focusing. cDNA clones will be isolated using the proposed polyclonal and monoclonal antibodies in order to determine ..
  27. BETA GALACTOSIDASES IN GLOBOID CELL LEUKODYSTROPHY
    Thaddeus Kurczynski; Fiscal Year: 1980
    ..Various enzyme fraction procedures, especially isoelectric focusing will be utilized...
  28. DEVELOPING T CELLS CLONES CYTOTOXIC TO SARCOMAS
    Michael Mastrangelo; Fiscal Year: 1990
    ..HLA-A2 subtypes responsible for target cell recognition will be defined using isoelectric focusing. The role of concordance at other HLA loci in the generation of specifically cytotoxic T cells will be ..
  29. STUDIES OF IGE DEPENDENT HISTAMINE RELEASING FACTORS
    Susan MacDonald; Fiscal Year: 1991
    ..Purification will be extended from ion-exchange and Sephadex chromatography to isoelectric focusing, polyacrylamide and SDS gel electrophoresis...
  30. STRUCTURE AND FUNCTION OF BACTERIORHODOPSIN
    Robert Stroud; Fiscal Year: 1992
    ..be solubilized in Triton X-100, purified in a sterol detergent by size exclusion, heterogeneity removed by isoelectric focusing in nonyl glucoside, and crystallized in a fourth detergent...
  31. CHARACTERIZATION OF RNA POL II TRANSCRIPTION PROTEINS
    Gyanendra Kumar; Fiscal Year: 1991
    ..proteins, after radiolabel transfer from the DNA, would then be identified by gel electrophoresis, isoelectric focusing and by tryptic mapping analysis...
  32. Top-Down Mass Spectrometry of Salivary Fluids for Cancer Assessment
    Joseph Loo; Fiscal Year: 2007
    ..Three methods will be investigated: separation of proteins by their hydrophobicity via HPLC; solution isoelectric focusing to partition the complex mixture into 5-7 separate sub-samples on the basis of their pl; affinity selection ..
  33. VARIABLE REGIONS OF BICLONAL IMMUNOGLOBULINS
    Robert Krueger; Fiscal Year: 1980
    ..With the triclonal proteins, we will determine if the three kappa-chains are identical by isoelectric focusing, mapping of iodinated tryptic and chymotryptic peptides and N-terminal amino acid sequence...
  34. INDUCTION AND MODE OF ACTION OF HEPATIC DRUG OXIDASE
    John Schenkman; Fiscal Year: 1992
    ..The purified proteins are characterized by NH2-terminal partialamino acid sequence, 2D-isoelectric focusing SDS-PAGE electrophoretograms, as well as by metabolite patterns with endogenous substrates like steriod ..
  35. INTRAOCULAR IMMUNOREGULATION AND IMMUNE COMPLEX DISEASE
    JOHN WALDREP; Fiscal Year: 1991
    ..The charge and quantity of Ig class/subclasses will be determined by ELISA, Immunoblotting, and Isoelectric Focusing of ocular tissue eluates...
  36. NEUROSPORA DNA REPAIR
    Alice Schroeder; Fiscal Year: 1980
    ..While these activities are separable on Hydroxyapatite chromatography and on isoelectric focusing, they are very similar in salt, Mg ions and pH optima and show identical chromatographic behavior on ..
  37. OVARIAN CANCER PROTEOME VIA TISSUE MICRODISSECTION AND GEMINI TECHNOLOGIES
    Satya Saxena; Fiscal Year: 2009
    ..techniques, including 2-D PAGE, multidimensional liquid chromatography systems, and gel and gel-free isoelectric focusing approaches such as chromatofocusing, immobilized pH membranes, Rotofor, and free-flow electrophoresis, are ..
  38. CHARACTERIZATION OF DISEASE-CAUSING ANTIBODIES IN EAMG
    Keith Krolick; Fiscal Year: 1993
    ..isolated with the use of specialized anti-idiotypic probes combined with methods of separation involving isoelectric focusing. Confirmation of myasthenogenicity will be performed by intravenous transfer od subseted antibodies into ..
  39. PROSTATIC GLUTATHIONE S-EPOXIDE TRANSFERASES
    CARL SCHAFFNER; Fiscal Year: 1991
    ..Enzyme homogeneity as well as pI values will be determined by isoelectric focusing (IEF)...
  40. OUTER MEMBRANE PERMEABILITY OF PSEUDOMONAS CEPACIA
    Jane Burns; Fiscal Year: 1992
    ..Next, the biochemical properties of each protein will be determined by gel electrophoresis, isoelectric focusing, peptide mapping and amino acid analysis...
  41. IMMUNOLOGICAL ASPECTS OF PREGNANCY IN HUMANS
    Thomas Gill; Fiscal Year: 1991
    ..Pa, 213, A/a and F/a isolated from trophoblast cells and from lymphocytes will be compared by SDS- PAGE, isoelectric focusing, sequential immunoprecipitation, peptide mapping and carbohydrate analysis...
  42. Development of Dynamic Isoelectric Focusing for Cancer Proteomics
    Luke Tolley; Fiscal Year: 2007
    ..Capillary isoelectric focusing (cIEF) has shown great promise in this area with a peak capacity in excess of 1400...
  43. GENE VARIATION IN NATURAL POPULATIONS
    Satya Prakash; Fiscal Year: 1980
    ..gel electrophoresis of proteins and enzymes in gels of different concentrations, different pH's and by isoelectric focusing. Polymorphism due to different number of cysteine residues with free sulfhydryls will be studied by ..
  44. CYCLIC AMP AND THE SECRETION OF LUTEINIZING HORMONE
    William Evans; Fiscal Year: 1990
    ..and biological properties of LH basally and in response to cyclic AMP will be determined by isoelectric focusing and bioassay...
  45. Accurate Early Detection of Preeclampsia
    BRADLEY ARBOGAST; Fiscal Year: 2001
    ..Preliminary results using an isoelectric focusing (IEF) assay demonstrated that levels of a subfraction of serum albumin could be used to accurately predict ..
  46. DIVERSITY OF HUMAN ANTIBODY TO H. INFLUENZAE B CAPSULE
    George Siber; Fiscal Year: 1991
    ..The technique of isoelectric focusing will be used to analyze the overall diversity of the human antibody to Hib capsular polysaccharide before ..
  47. STUDIES OF A NEWLY DISCOVERED HUMAN IGG FC RECEPTOR
    R Looney; Fiscal Year: 1990
    ..3. We will use isoelectric focusing gels and peptide mapping to see if the B cell receptor is closely related to the receptor recognized by IV...
  48. CHARACTERIZATION/ISOLATION--BIOCALCIFICATION INHIBITORS
    SYBIL BURGESS; Fiscal Year: 1992
    ..Polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing (IEF) will be conducted on cuticle tissue from all crab molt stages...
  49. Proteomics of Progression in MCF10 Xenograft Model
    Fred Miller; Fiscal Year: 2005
    ..The method will be combined with a first dimension separation using liquid phase isoelectric focusing (IEF) to prefractionate proteins according to pI...
  50. SUPPRESSION OF HIV REPLICATION BY CD8+ CELLS
    Jay Levy; Fiscal Year: 1999
    ..The approaches include standard protein purification methods (i.e., column chromatography, isoelectric focusing, and gel electrophoresis) using an algorithm that removes albumin from the serum-free medium and permits ..
  51. FINAL EXPRESSION OF BETA-GLUCURONIDASE IN DROSOPHILA
    Daniel Hartl; Fiscal Year: 1980
    ..first years have established; (1) Drosophila has three or possibly four forms of the enzyme as observed in isoelectric focusing gels, (2) the principal activity is due to a soluble form of the enzyme of molecular weight 290,000 daltons,..
  52. HIGH SENSITIVITY PEPTIDE SEQUENCING BY CE/MS AND CE/MS-M
    CHARLES EDMONDS; Fiscal Year: 1990
    ..zone electrophoresis, electrophoresis in gel filled capillaries, capillary isotachophoresis or capillary isoelectric focusing with dynamic detection Dy electrospray ionization mass spectrometry...
  53. STRUCTURE ELUCIDATION OF HUMAN TRANSFER FACTOR
    G Daves; Fiscal Year: 1980
    ..to a selected antigen (KLH) is only apparent after immunization, (b) TF can be purified by gel filtration, isoelectric focusing (IEF), and reverse phase, high pressure liquid chromatography (HPLC), (c) the purified TF is not ..
  54. CHARACTERIZATION OF CHOLINERGIC NEUROTROPHIC FACTOR
    BRADLEY WISE; Fiscal Year: 1991
    ..hydrophobic interaction chromatography, ion-exchange chromatography, gel filtration chromatography, isoelectric focusing, and high performance liquid chromatography will be used to purify the neurotrophic protein present in ..
  55. GENETIC CONTROL OF IMMUNOGLOBULIN LIGHT CHAINS
    David Gibson; Fiscal Year: 1980
    Analysis of immunoglobulin light chains by gel isoelectric focusing led to the discovery of genetic polymorphism in certain inbred strains of mice...
  56. QUANTITATIVE ASSESSMENT OF SUSTAINED ALCOHOL USAGE
    SAMAR MAKHLOUF; Fiscal Year: 1990
    ..The pI 5.7 transferrin isoform can be detected by isoelectric focusing and its quantities have been shown to directly correlate with the amount of alcohol intake by an individual ..
  57. NEW APPROACH FOR RAPID MAPPING OF PROTEIN VARIATIONS
    Philip Andrews; Fiscal Year: 2000
    ..This method works well with isoelectric focusing (IEF), native, and SDS gels, and has been applied to 3-D analysis of complex protein mixtures with ..
  58. Ultrasensitive Approaches for Complex Animal Proteomics
    Yan Li; Fiscal Year: 2005
    The R43 Phase of this project aims to develop and demonstrate a capillary isoelectric focusing-based multidimensional separation platform, capable of providing significant analyte concentration and extremely high resolving power for ..
  59. Chaperone rich cell lysates (CRCL)Natural adjuvants and*
    Emmanuel Katsanis; Fiscal Year: 2004
    ..method that efficiently enriches for multiple chaperone complexes from tumor lysates using free solution isoelectric focusing. We have documented that chaperone rich cell lysate (CRCL) vaccination is more effective than immunization ..
  60. CLONAL EVOLUTION OF HUMAN MALIGNANT LYMPHOMAS
    RICHARD RUDDERS; Fiscal Year: 1991
    ..against the idiotypic portions of the Ig molecule as well as to detect subtle Ig structural changes by isoelectric focusing in gels...
  61. ROLE OF GLUCOSE TRANSPORT PROTEINS IN DIABETES MELLITUS
    W Garvey; Fiscal Year: 1992
    ..To test for structural abnormalities, we will combine SDS-PAGE and isoelectric focusing to determine the MW, charge heterogeneity, and extent of post-translational glycosylation of photolabeled ..
  62. An Automated 2D-CAE System Interfaced to MALDI MS
    GARY LOGE; Fiscal Year: 2006
    ..Innovations include a scheme to couple isoelectric focusing (CIEF) in a glass micro-fluidic chip channel with gel electrophoresis (CGE) in a capillary array with ..
  63. 2D PROTEIN ANALYSIS USING MICROCHIP MASS SPECTROMETRY
    Cheng Lee; Fiscal Year: 2000
    ..analogy to a 2-D analysis system, fundamental studies on the fabrication of 3-D MEMS and the performance of isoelectric focusing in the microchannel establish the basis for the first separation mechanism...