- Regulation of microfilament organization by Kaposi sarcoma-associated herpes virus-cyclin.CDK6 phosphorylation of caldesmon
Maria Emanuela Cuomo
Cancer Research UK Centre for Cell and Molecular Biology, Chester Beatty Laboratories, The Institute of Cancer Research, 237 Fulham Road, SW3 6JB London, United Kingdom
J Biol Chem 280:35844-58. 2005
..These results establish a novel link between KSHV infection and the regulation of the actin cytoskeleton...
- A self-assembling process in articular cartilage tissue engineering
Jerry C Hu
Department of Bioengineering, Rice University, Houston, Texas 77005, USA
Tissue Eng 12:969-79. 2006
..IHC and gel electrophoresis showed collagen type II production and absence of collagen type I. More importantly, self-assembled constructs reached well over one third the stiffness of native tissue...
- Novel application of S-nitrosoglutathione-Sepharose to identify proteins that are potential targets for S-nitrosoglutathione-induced mixed-disulphide formation
Department of Estructura y Función de Proteínas, Centro de Investigaciones Biologicas, Instituto Reina Sofía de Investigaciones Nefrológicas, Consejo Superior de Investigaciones Cientificas, Velazquez 144, E 28006 Madrid, Spain
Biochem J 349:567-78. 2000
..Here, we show that GSNO-Sepharose mimicks site-specific S-glutathionylation of the transcription factors c-Jun and p50 by free GSNO in vitro...
- Purification and characterization of three toxins and two agglutinins from Abrus precatorius seed by using lactamyl-Sepharose affinity chromatography
Department of Biochemistry, Indian Institute of Science, Bangalore
Anal Biochem 194:101-9. 1991
..II, and -III, and two agglutinins, APA-I and -II, were purified from the seeds of Abrus precatorius by lactamyl-Sepharose affinity chromatography followed by gel filtration and DEAE-Sephacel column chromatography...
- Quantification of mRNA using real-time PCR and Western blot analysis of MAPK events in chondrocyte/agarose constructs
David A Lee
School of Engineering and Materials Science, Queen Mary University of London, London, UK
Methods Mol Biol 695:77-97. 2011
..Ultimately, development of methods used to analyse mechano-sensitive pathways will provide important information for the identification of appropriate pharmacological and physiotherapeutic agents for the treatment of osteoarthritis...
- Integrin-mediated mechanotransduction in IL-1 beta stimulated chondrocytes
T T Chowdhury
Medical Engineering Division and IRC in Biomedical Materials, Department of Engineering, Queen Mary, University of London, Mile End Road, London, E1 4NS, UK
Biomech Model Mechanobiol 5:192-201. 2006
..The non-binding control peptide, GRADSP, had no effect. These data clearly demonstrate that the metabolic response of the chondrocytes to dynamic compression in the presence and absence of IL-1 beta, are integrin mediated...
- Selection of thiol- and disulfide-containing proteins of Escherichia coli on activated thiol-Sepharose
Proteomics Research Group, Department of Biochemistry, University College Cork, Cork, Ireland
Anal Biochem 398:245-53. 2010
Activated thiol-Sepharose (ATS) facilitates selection of thiol-containing proteins...
- Dynamic compression inhibits the synthesis of nitric oxide and PGE(2) by IL-1beta-stimulated chondrocytes cultured in agarose constructs
T T Chowdhury
IRC in Biomedical Materials and Medical Engineering Division, Department of Engineering, Queen Mary, University of London, Mile End Road, London, E1 4NS, United Kingdom
Biochem Biophys Res Commun 285:1168-74. 2001
..The inhibition of (*)NO and PGE(2) by dynamic compression is a finding of major significance that could contribute to the development of novel strategies for the treatment of cartilage-degenerative disorders...
- Hydrophobic interaction chromatography of homo-oligonucleotides on derivatized Sepharose CL-6B. Application of the solvophobic theory
M M Diogo
Centro de Engenharia Biologica e Quimica, Instituto Superior Tecnico, Lisboa, Portugal
J Chromatogr A 944:119-28. 2002
..interaction chromatography of homo-deoxyoligonucleotides polyA, polyT and polyU with sizes up to 30 bases on a Sepharose gel derivatized with the 1,4-butanediol diglycidyl ether...
- Spatial and temporal development of chondrocyte-seeded agarose constructs in free-swelling and dynamically loaded cultures
Terri Ann N Kelly
Cellular Engineering Laboratory, Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA
J Biomech 39:1489-97. 2006
- Removal of endogenous retrovirus-like particles from CHO-cell derived products using Q sepharose fast flow chromatography
Daniel M Strauss
Process Research and Development, Late Stage Purification, Genentech Inc, One DNA Way, South San Francisco, CA 94080, USA
Biotechnol Prog 25:1194-7. 2009
..We have determined the ability of a typical Q sepharose process to remove actual CHO RVLP impurities...
- Modulation of penicillin acylase properties via immobilization techniques: one-pot chemoenzymatic synthesis of Cephamandole from Cephalosporin C
Pharmaceutical Biocatalysis Laboratories, Dipartimento di Chimica Farmaceutica, Universita degli Studi di Pavia, Via Taramelli 12, 27100, Pavia, Italy
Bioorg Med Chem Lett 11:2429-32. 2001
- Nitric oxide-activated glutathione Sepharose
Department of Immunology and Oncology, Centro Nacional de Biotecnologia, Campus of Cantoblanco, 28049 Madrid, Spain
Methods Enzymol 359:245-55. 2002
- Molecular analysis of chondrocytes cultured in agarose in response to dynamic compression
UMR 5086, CNRS, Universite de Lyon, IFR 128, IBCP, Institut de Biologie et Chimie des Proteines, 7 passage du Vercors F 69367 Lyon FRANCE
BMC Biotechnol 8:71. 2008
..The mouse model is of particular interest because of the availability of a large choice of bio-molecular tools suitable to study it, as well as genetically modified mice...
- Antibody purification: affinity chromatography - protein A and protein G Sepharose
Ana Cristina Grodzki
Receptors and Signal Transduction Section, Oral Infection and Immunity Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, USA
Methods Mol Biol 588:33-41. 2010
..In the Subheading 3 there are also protocols for affinity purification using a specific ligand coupled to commercial matrices like CNBr- Sepharose 4-B and Affigel.
- Proteomic characterization of protein phosphatase complexes of the mammalian nucleus
Hue T Tran
Department of Biological Sciences, University of Calgary, Calgary, Alberta T2N 1N4, Canada
Mol Cell Proteomics 3:257-65. 2004
..Microcystin-Sepharose chromatography and mass spectrometry were utilized to affinity purify and identify protein phosphatase-..
- Measurement of the deformation of isolated chondrocytes in agarose subjected to cyclic compression
M M Knight
IRC in Biomedical Materials, Queen Mary and Westfield College, London, UK
Med Eng Phys 20:684-8. 1998
..72), although during static compression the cell deformation remained constant. This system may be used to examine cellular events under a range of dynamic mechanical stimuli...
- Dynamic compression counteracts IL-1beta induced inducible nitric oxide synthase and cyclo-oxygenase-2 expression in chondrocyte/agarose constructs
T T Chowdhury
School of Engineering and Materials Science, Queen Mary, University of London, Mile End Road, London, E1 4NS, UK
Arthritis Res Ther 10:R35. 2008
- A protocol for separation and isolation of small and/or large DNA fragments with high yield using CL4B Sepharose
Department of Nuclear Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, 10 Center Drive, Bethesda, MD 20892, USA
Anal Biochem 317:284-7. 2003
- Efficient removal of PCR inhibitors using agarose-embedded DNA preparations
Laboratoire de Biologie Cellulaire BC4, URA CNRS 2227, Universite Paris Sud, Bâtiment 444, 91405 Orsay Cedex, France
Nucleic Acids Res 26:3309-10. 1998
- Single-step method for purification of Shiga toxin-1 B subunit using receptor-mediated affinity chromatography by globotriaosylceramide-conjugated octyl sepharose CL-4B
Department of Pathology, National Children's Medical Research Center, Taishido, Tokyo, Japan
Protein Expr Purif 22:267-75. 2001
..using receptor-mediated affinity chromatography, in which Gb3Cer (globotriaosylceramide) was conjugated to octyl Sepharose CL-4B as a carrier...
- Bioaffinity immobilization of tannase from Aspergillus niger on concanavalin A-Sepharose CL-4B
Chemistry Department, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi, India
Biotechnol Appl Biochem 35:165-9. 2002
Tannase from Aspergillus niger van Teighem was immobilized on concanavalin A-Sepharose via bioaffinity interaction. The immobilized enzyme showed a pH optimum similar to that of the free enzyme...
- Purinergic pathway suppresses the release of .NO and stimulates proteoglycan synthesis in chondrocyte/agarose constructs subjected to dynamic compression
T T Chowdhury
Medical Engineering Division, Department of Engineering, Queen Mary, University of London, Mile End Road, London, UK
J Cell Physiol 209:845-53. 2006
..The current study demonstrates for the first time the importance of a purinergic pathway in mediating the metabolic response to dynamic compression and suppressing an inflammatory effect...
- Improved in vitro development of OPU-derived bovine (Bos taurus) embryos by group culture with agarose-embedded helper embryos
E M Senatore
ARSIAL Centro Regionale per la Zootecnia, Rome, Italy
Theriogenology 74:1643-51. 2010
..1%, higher than that in the 3 + 0 group (11.8%, P < 0.05). In conclusion, limited numbers of OPU/IVF oocytes had competent development when cultured with helpers (embedded in agarose to provide physical separation)...
- Rapid purification and characterization of nucleoside diphosphate kinase isoforms using ATP-sepharose affinity column chromatography
S Y Kim
College of Pharmacy, Ewha Womans University, Seoul, Korea
Mol Cells 7:630-4. 1997
..with pET3C expression plasmids containing nm23-H1 or nm23-H2, were purified in one step to homogeneity using ATP-sepharose affinity column chromatography...
- Improved isolation of undersampled bacteriophages: finding of distant terminase genes
Department of Biochemistry, The University of Texas Health Science Center, San Antonio, TX 78229 3900, USA
Virology 329:412-24. 2004
..Some had genomes with extremely distant relationships to those of other bacteriophages, based on a tree built from the large terminase genes. These methods find novel genomes by rapidly isolating and screening diverse bacteriophages...
- Dynamic compression counteracts IL-1 beta-induced release of nitric oxide and PGE2 by superficial zone chondrocytes cultured in agarose constructs
T T Chowdhury
Medical Engineering Division and IRC in Biomedical Materials, Department of Engineering, Queen Mary, University of London, Mile End Road, London, UK
Osteoarthritis Cartilage 11:688-96. 2003
..To examine the effect of IL-1 beta-induced *NO and PGE(2)release by stimulated superficial and deep chondrocyte/agarose constructs subjected to mechanical compression...
- Isolation of soybean trypsin inhibitors by affinity chromatography on anhydrotrypsin-Sepharose 4B
Rowett Research Institute, Bucksburn, Aberdeen, Scotland, United Kingdom
Anal Biochem 172:108-12. 1988
..Inactive material was removed by affinity chromatography on trypsin inhibitor-Sepharose 4B and the purified anhydrotrypsin with full binding capacity for trypsin inhibitors was coupled to cyanogen ..
- Pulsatile dynamic stiffness of cartilage-like materials and use of agarose gels to validate mechanical methods and models
P Scandiucci de Freitas
Laboratory for Orthopedic Biomechanics, LOB, University of Basel, c o Biozentrum Pharmazentrum, Klingelbergstrasse 50 70, 4056 Basel, Switzerland
J Biomed Mater Res B Appl Biomater 78:347-57. 2006
..Also, agarose is hygroscopic, leading to unintended variations in gel composition. Agarose gels are problematic materials for validation or comparison of cartilage mechanical test methods and models...
- Purification of Shiga-like toxin 1 by pigeon egg white glycoproteins immobilized on Sepharose gels
Kitasato Institute for Life Sciences, Kitasato University, 5 9 1 Shirokane, Minato ku, Tokyo 108 8641, Japan
Anal Biochem 311:50-6. 2002
..Pigeon ovalbumin, ovomucoid, or the whole egg white were immobilized on periodate-oxidized Sepharose CL-6B gels by reductive amination...
- Mechanical loading of chondrocytes embedded in 3D constructs: in vitro methods for assessment of morphological and metabolic response to compressive strain
David A Lee
IRC in Biomedical Materials and Medical Engineering Division, Department of Engineering, Queen Mary College, University of London, London, UK
Methods Mol Med 100:307-24. 2004
..The chapter thus provides an introduction to the experimental techniques used to examine chondrocyte mechanotransduction and downstream cell function...
- Inclusion of S-sepharose beads in the culture medium significantly improves recovery of secreted rBPI(21) from transfected CHO-K1 cells
A H Horwitz
XOMA US LLC, 1545 17th Street, Santa Monica, California, 90404, USA
Protein Expr Purif 18:77-85. 2000
..To address these issues, we developed a novel process whereby sterile S-Sepharose beads were added directly to the cell culture medium...
- Temporal regulation of chondrocyte metabolism in agarose constructs subjected to dynamic compression
Tina T Chowdhury
Medical Engineering Division, Department of Engineering, Queen Mary, University of London, Mile End Road, London E1 4NS, UK
Arch Biochem Biophys 417:105-11. 2003
..The uncoupled nature of the metabolic response determined in this study suggests that mechanical conditioning regimes may be fine tuned to selectively stimulate key metabolic parameters of relevance to cartilage tissue engineering...
- Purification of elongation factors EF-Tu and EF-G from Escherichia coli by covalent chromatography on thiol-sepharose
T D Caldas
Biochimie Génétique, Institut Jacques Monod, Universite Paris 7, 2 place Jussieu, Paris Cedex 05, 75251, France
Protein Expr Purif 14:65-70. 1998
..We took advantage of this property to design a purification protocol based on thiol-Sepharose chromatography, a method involving thiol-disulfide interchange between protein thiol groups and the glutathione-..
- The influence of elaborated pericellular matrix on the deformation of isolated articular chondrocytes cultured in agarose
M M Knight
IRC in Biomedical Materials, Queen Mary and Westfield College, Mile End Road, London E1 4NS, UK
Biochim Biophys Acta 1405:67-77. 1998
- Heparin elution of transcription factors from DNA-Sepharose columns
Department of Biochemistry, University of Tennessee, Memphis 38163, USA
J Chromatogr A 848:131-8. 1999
A novel method using heparin for eluting transcription factors from DNA-Sepharose columns was characterized...
- Hydrostatic pressure modulates proteoglycan metabolism in chondrocytes seeded in agarose
Keio University, Tokyo, Japan
Arthritis Rheum 48:2865-72. 2003
..To investigate the effect of isolated hydrostatic pressure on proteoglycan metabolism in chondrocytes...
- Sepharose-unbinding ricin E as a source for ricin A chain immunotoxin
B H Woo
Drug Targeting Laboratory, College of Pharmacy, Sungkyunkwan University, 300 Chonchon-dong, Jangan-ku, 440-746, Suwon City, South Korea
J Immunol Methods 249:91-8. 2001
To evaluate the Sepharose-unbinding ricin E as a preference source material for ricin A chain (RTA) in immunotoxin studies, RTA of ricin E (RTA(E)) was characterized and compared with RTA of the Sepharose-binding ricin D (RTA(D))...
- Investigating conversion of mechanical force into biochemical signaling in three-dimensional chondrocyte cultures
UMR5086, CNRS, IFR128, IBCP Institut de Biologie et Chimie des Protéines, Universite de Lyon, Lyon, France
Nat Protoc 4:928-38. 2009
..The protocols can be carried out with a limited number of mouse embryos or newborns and this point is particularly important regarding genetically modified mice...
- Dynamic compression inhibits fibronectin fragment induced iNOS and COX-2 expression in chondrocyte/agarose constructs
S P Raveenthiran
School of Engineering and Materials Science, Queen Mary, University of London, London, E1 4NS, UK
Biomech Model Mechanobiol 8:273-83. 2009
..The combination of mechanical and pharmacological interventions makes this study a powerful tool to examine further the interactions of biomechanics and cell signalling in osteoarthritis...
- Effects of dynamic compressive loading on chondrocyte biosynthesis in self-assembling peptide scaffolds
John D Kisiday
Biological Engineering Division, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA
J Biomech 37:595-604. 2004
..Taken together, these results demonstrate the potential of dynamic compression for stimulating PG synthesis and accumulation for applications to in vitro culture of tissue engineered constructs prior to implantation...
- Templated agarose scaffolds support linear axonal regeneration
Department of Bioengineering, University of California San Diego, La Jolla, California 92093, USA
Tissue Eng 12:2777-87. 2006
..The templating process can be useful in fabricating nerve guidance scaffolds for both central and peripheral nerve injuries, or any materials application requiring a precise array of linearly oriented channels...
- Synthesis and use of the protein phosphatase affinity matrices microcystin-sepharose and microcystin-biotin-sepharose
Greg B G Moorhead
Department of Biological Sciences, University of Calgary, Alberta, Canada
Methods Mol Biol 365:39-45. 2007
..Here, we describe the procedure for the synthesis and use of microcystin-Sepharose and microcystin-biotin-Sepharose.
- Dynamic compression counteracts IL-1beta induced iNOS and COX-2 activity by human chondrocytes cultured in agarose constructs
Tina T Chowdhury
Medical Engineering Division and IRC in Biomedical Materials, Department of Engineering, Queen Mary, University of London, Mile End Road, London, E1 4NS, UK
Biorheology 43:413-29. 2006
..These data indicate interactions exist between the NOS and COX pathways, a finding which will provide new insights in the development of pharmacological or biophysical treatments for cartilage disorders such as OA...
- Comparative studies on discrete and concatemeric DNA-sepharose columns for purification of transcription factors
Department of Biochemistry, University of Tennessee, Memphis 38163, USA
J Chromatogr A 917:43-53. 2001
..Our study shows concatemeric columns do not have any distinct advantage for the three different transcription factors we studied including Sp1, the original justification for the concatemeric approach...
- Protein A Sepharose immunoadsorption can restore the efficacy of platelet concentrates in patients with Glanzmann's thrombasthenia and anti-glycoprotein IIb-IIIa antibodies
Laboratoire d Hematologie, Centre de Traitement de l Hémophilie, Assistance Publique Hôpitaux de Paris and Faculté de Médecine Paris XI, Le Kremlin Bicetre, Paris, France
Br J Haematol 119:991-7. 2002
..We used immunoadsorption on protein A Sepharose (IA-PA), which has been shown to be efficient in decreasing the titre of antibodies in several immune diseases, ..
- Enzymatically inactive trans-sialidase from Trypanosoma cruzi binds sialyl and beta-galactopyranosyl residues in a sequential ordered mechanism
Adriane R Todeschini
Instituto de Biofisica Carlos Chagas Filho, Centro de Ciências da Saúde Bloco G, Universidade Federal do Rio de Janeiro, 21 944970, Cidade Universitaria, Ilha do Fundao, Rio de Janeiro, Brasil
J Biol Chem 279:5323-8. 2004
..cruzi/host cell interaction and could shed more light into the catalytic mechanism of the sialic acid transfer reaction of enzymatically active trans-sialidase...
- Stabilization of pancreatic ribonuclease A by immobilization on Sepharose-linked antibodies that recognize the labile region of the enzyme
Interdisciplinary Biotechnology Unit, Faculty of Life Sciences, Aligarh Muslim University, India
Biochim Biophys Acta 1548:114-20. 2001
..RNase immobilized on Sepharose support precoupled either with the antipeptide immunoglobulin (IgG) or anti-RNase IgG proved to be more ..
- Boronate-containing copolymers: polyelectrolyte properties and sugar-specific interaction with agarose gel
Marina V Kuzimenkova
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P O Box 124, SE 22100, Lund, Sweden
Macromol Biosci 6:170-8. 2006
..DMAA), and N-isopropylacrylamide (NIPAM) were found to adsorb on cross-linked agarose gel (Sepharose CL-6B) in the pH range from 7.5-9.2, due to specific boronate-sugar interactions...
- Activation of the p70 S6 kinase and phosphorylation of the 4E-BP1 repressor of mRNA translation by type I interferons
Robert H Lurie Comprehensive Cancer Center and Division of Hematology Oncology, Northwestern University Medical School and Lakeside Veterans Administration Medical Center, Chicago, Illinois 60611, USA
J Biol Chem 278:27772-80. 2003
..Altogether, our data establish that the Type I IFN receptor-activated PI 3'-kinase pathway mediates activation of the p70 S6 kinase and inactivation of 4E-BP1, to regulate mRNA translation and induction of Type I IFN responses...
- Monosize poly(glycidyl methacrylate) beads for dye-affinity purification of lysozyme
Evrim Banu Altıntas
Department of Chemistry, Biochemistry Division, Hacettepe University, Ankara, Turkey
Int J Biol Macromol 38:99-106. 2006
..The purity of the eluted lysozyme was analyzed by SDS-PAGE and found to be 88% with recovery about 79%. The specific activity of the eluted lysozyme was high as 43,600 U/mg...
- Characterization of two heparan sulphate-binding sites in the mycobacterial adhesin Hlp
Michelle I Portugal
Laboratório de Microbiologia Celular, Instituto Oswaldo Cruz, FIOCRUZ, Av, Brasil 4365, Manguinhos, Rio de Janeiro, RJ 21045 900, Brazil
BMC Microbiol 8:75. 2008
..In the present study, nuclear magnetic resonance (NMR) was used to map the binding site(s) of Hlp to heparan sulfate and identify the nature of the amino acid residues directly involved in this interaction...
- Identification and characterization of beta-D-galactosyl-transferase in chick corneas
Radiochemistry Section, Faculty of Pharmacy, Meijo University, Nagoya, Japan
Jpn J Ophthalmol 46:607-15. 2002
..To purify beta-galactosyltransferase, which is involved in the biosynthesis of keratan sulfate, from 2-day-old chick corneas...
- Protein-protein interaction assays: eliminating false positive interactions
Tuan N Nguyen
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, USA
Nat Methods 3:135-9. 2006
- Biosensor-based micro-affinity purification for the proteomic analysis of protein complexes
The Ludwig Institute for Cancer Research, Melbourne Tumor Biology Branch, PO Box 2008, Royal Melbourne Hospital, Victoria 3050, Melbourne, Australia
J Proteome Res 4:1646-56. 2005
..The results obtained using the SPU were compared with similar experiments performed using Sepharose beads.
- Screening for beta-poly(L-malate) binding proteins by affinity chromatography
Institut fur Biophysik und Physikalische Biochemie, Universitat Regensburg, D 93040 Regensburg, Germany
Biochem Biophys Res Commun 341:1119-27. 2006
..We have developed an affinity chromatography employing 1,6-diamino-n-hexane-Sepharose-coupled poly(malic acid) to identify such proteins in cellular extracts of Physarum polycephalum...
- 4-N-trimethylaminobutyraldehyde dehydrogenase: purification and characterization of an enzyme from Pseudomonas sp. 13CM
United Graduate School of Agricultural Sciences, Tottori University, Koyama, Tottori, Japan
Biosci Biotechnol Biochem 72:155-62. 2008
..The K(m) values for 4-N-trimethylaminobutyraldehyde, 4-dimethylaminobutyraldehyde, and NAD(+) were 7.4, 51, and 125 microM respectively. The enzyme was inhibited by SH reagents, and by heavy metal ions...
- Novel hydrophobic interaction chromatography matrix for specific isolation and simple elution of immunoglobulins (A, G, and M) from porcine serum
Centro de Investigación en Alimentación y Desarrollo A C Coordinación de Ciencia de los Alimentos, Apartado Postal 1735, Hermosillo, Sonora 83000, Mexico
J Chromatogr A 1122:28-34. 2006
A new, highly acetylated agarose matrix (HA-Sepharose) was synthesized and used as a hydrophobic interaction chromatography (HIC) medium to specifically isolate immunoglobulins (Igs) from porcine serum...
- FCP1 phosphorylation by casein kinase 2 enhances binding to TFIIF and RNA polymerase II carboxyl-terminal domain phosphatase activity
UMR 8541 CNRS, Genetique Moleculaire, Ecole Normale Superieure, 46 rue d Ulm, 75230 Paris Cedex 05, France
J Biol Chem 277:36061-7. 2002
..These findings unravel a new mechanism regulating CTD phosphorylation and hence class II gene transcription...
- Induced binding of proteins by ammonium sulfate in affinity and ion-exchange column chromatography
Alliance Protein Laboratories, 3957 Corte Cancion, Thousand Oaks, CA 91360, USA
J Biochem Biophys Methods 70:493-8. 2007
..Blue-Sepharose is often used to remove bovine serum albumin (BSA) from samples, but when we applied BSA to Blue-Sepharose in 20 ..
- Chemical and physicochemical characterization of the sialic acid-specific lectin from Cepaea hortensis
Friedrich Schiller University of Jena, Institute of Medical Microbiology, Semmelweisstr 4, Germany
FEMS Microbiol Lett 214:61-8. 2002
..was isolated from the albumin glands of the garden snail Cepaea hortensis by affinity chromatography on fetuin-Sepharose following gel filtration on Superdex 200...
- Galactosyl-mimodye ligands for Pseudomonas fluorescens beta-galactose dehydrogenase
C F Mazitsos
Laboratory of Enzyme Technology, Department of Agricultural Biotechnology, Agricultural University of Athens, Greece
Eur J Biochem 269:5391-405. 2002
..Therefore, these new mimodyes suggested by computational analysis, are candidates for application in affinity labeling and structural studies as well as for purification of galactose dehydrogenase...
- A heat shock protein70 fusion protein with alpha1-antitrypsin in plasma of type 1 diabetic subjects
Department of Pharmacology and Anaesthesiology, University of Padua, Largo E Meneghetti 2, 35131 Padua, Italy
Biochem Biophys Res Commun 315:297-305. 2004
..The additional finding of circulating antibodies against the HSP70-alpha1AT protein supported its immunogenic properties with implications for diabetes and its complications...
- Identification of serine proteases from Leishmania braziliensis
Herbert L M Guedes
Laboratório de Bioquímica de Proteínas e Peptídeos, Departamento de Bioquimica e Biologia Molecular, Fundacao Oswaldo Cruz, Rio de Janeiro, RJ, Brazil
Z Naturforsch C 62:373-81. 2007
..amazonensis serine proteases, demonstrating a conservation of this enzymatic class in the Leishmania genus. This is the first study to report the purification of a serine protease from Leishmania braziliensis...
- Beta 1,4-galactosyltransferase (beta 4GalT)-IV is specific for GlcNAc 6-O-sulfate. Beta 4GalT-IV acts on keratan sulfate-related glycans and a precursor glycan of 6-sulfosialyl-Lewis X
Department of Biochemistry, Sasaki Institute, Kanda Surugadai 2 2, Tokyo 101 0062, Japan
J Biol Chem 278:9150-8. 2003
..These results suggested that beta4GalT-IV is a 6SGN-specific beta4GalT and may be involved in the biosynthesis of various glycoproteins carrying a 6-O-sulfated N-acetyllactosamine moiety...
- Nucleotide-mimetic synthetic ligands for DNA-recognizing enzymes One-step purification of Pfu DNA polymerase
Laboratory of Enzyme Technology, Department of Agricultural Biotechnology, Agricultural University of Athens, 75 Iera Odos Street, GR 11855 Athens, Greece
J Chromatogr A 1122:63-75. 2006
..Quality control tests showed that Pfu Pol purified on the B-motif-complementing ligand is free of nucleic acids and contaminating nuclease activities, therefore, suitable for experimental use...
- Purification and characterization of recombinant, human acid ceramidase. Catalytic reactions and interactions with acid sphingomyelinase
Department of Human Genetics, Mount Sinai School of Medicine, New York, New York 10029, USA
J Biol Chem 278:32978-86. 2003
..These studies provide new insights into acid ceramidase and the related lipid hydrolase, acid sphingomyelinase...
- Purification, characterization, and sequence analysis of two alpha-amylase isoforms from azuki bean, Vigna angularis, showing different affinity towards beta-cyclodextrin sepharose
San San Mar
Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo 060 8589, Japan
Biosci Biotechnol Biochem 67:1080-93. 2003
..VAAmy1 apparently had lower affinity towards a beta-cyclodextrin Sepharose column than VAAmy2. Molecular weights of VAAmy1 and VAAmy2 were estimated to be 47,000 and 44,000, respectively...
- ADAM 12 cleaves extracellular matrix proteins and correlates with cancer status and stage
Program in Vascular Biology and Department of Surgery, Children s Hospital, Boston, Massachusetts 02115, USA
J Biol Chem 279:51323-30. 2004
..We purified ADAM 12 from the urine of breast cancer patients via Q-Sepharose anion exchange and gelatin-Sepharose affinity chromatography followed by protein identification by matrix-..
- Characterization of growth factor-binding structures in heparin/heparan sulfate using an octasaccharide library
Institute for Molecular Science of Medicine, Aichi Medical University, Nagakute, Aichi 480 1195, Japan
J Biol Chem 279:12346-54. 2004
..Thus, the library may provide new reagents to specifically regulate bindings of the growth factors to HS...
- Purification of sequence-specific DNA-binding proteins by affinity chromatography
L A Kerrigan
University of California San Diego, La Jolla, California, USA
Curr Protoc Mol Biol . 2001
..An provides a method to couple DNA to commercially available CNBr-activated Sepharose, and a support protocol describes how to purify crude synthetic oligonucleotides by gel electrophoresis prior to ..
- Gentamicin binds to the lectin site of calreticulin and inhibits its chaperone activity
Department of Bioscience and Technology, Faculty of Science and Engineering, Ritsumeikan University, 1 1 1 Noji higashi, Kusatsu, Shiga 525 8577, Japan
Biochem Biophys Res Commun 323:281-7. 2004
..Experiments with domain mutants suggest that the lectin site of CRT is the main target for gentamicin binding and that binding of gentamicin to this site inhibits the chaperone activity of CRT...
- A model for the salt effect on adsorption equilibrium of basic protein to dye-ligand affinity adsorbent
Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, People s Republic of China
Biotechnol Prog 20:207-14. 2004
..describing the salt effect on adsorption equilibrium of a basic protein, lysozyme, to Cibacron Blue 3GA-modified Sepharose CL-6B (CB-Sepharose) has been developed...
- MeCP2 behaves as an elongated monomer that does not stably associate with the Sin3a chromatin remodeling complex
Robert J Klose
Wellcome Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3JR, Scotland, United Kingdom
J Biol Chem 279:46490-6. 2004
..Our findings indicate the MeCP2 is not an obligate component of the Sin3a corepressor complex and may therefore engage a more diverse range of cofactors for repressive function...
- Chemically selective displacers for high-resolution protein separations in ion-exchange systems: effect of displacer-protein interactions
Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Ricketts Building, 110 8th Street, Troy, New York 12180, USA
Anal Chem 80:3357-64. 2008
..The ability to exploit protein displacer binding in concert with appropriate displacer resin affinities opens up new possibilities for creating selective displacement systems...
- A binding site for highly sulfated heparan sulfate is identified in the N terminus of the circumsporozoite protein: significance for malarial sporozoite attachment to hepatocytes
John B Ancsin
Department of Pathology and Molecular Medicine, Queen s University and the Syl and Molly Apps Research Center, Kingston General Hospital, Kingston, Ontario Canada, K7L 3N6
J Biol Chem 279:21824-32. 2004
..high affinity heparin/HS binding activity and the ability to block the binding of recombinant CSP to heparin-Sepharose 4B. Analysis by isothermal titration calorimetry revealed that region I-plus has a binding constant of K(d) = 5...
- A protein-protein binding assay using coated microtitre plates: increased throughput, reproducibility and speed compared to bead-based assays
Tim J Craig
Physiological Laboratory, University of Liverpool, Crown Street, PO Box 147, Liverpool L69 3BX, UK
J Biochem Biophys Methods 60:49-60. 2004
..we have modified this form of binding assay, using glutathione-coated 96-well plates rather than glutathione-Sepharose beads to bind the primary bait protein...
- Rational affinity purification of native Streptomyces family 10 xylanase
Department of Material and Biological Chemistry, Faculty of Science, Yamagata University, Yamagata 990 8560, Japan
J Biotechnol 110:137-42. 2004
..Lactosyl-Sepharose was prepared and tested as an affinity matrix...
- Cloning, characterization, and structural analysis of a C-type lectin from Bothrops insularis (BiL) venom
Rede Proteômica do Rio de Janeiro and Laboratório de Hemostase e Venenos LabHemoVen, Departamento de Bioquímica Médica ICB, Universidade Federal do Rio de Janeiro, CEP 21941 590, Rio de Janeiro RJ, Brazil
Arch Biochem Biophys 432:1-11. 2004
..BiL shows similarity to other C-type lectin family members. Modeling studies provide insights into BiL dimeric structure and its structural determinants for carbohydrate and calcium binding...
- The small heat shock protein IbpA of Escherichia coli cooperates with IbpB in stabilization of thermally aggregated proteins in a disaggregation competent state
Department of Molecular and Cellular Biology, Intercollegiate Faculty of Biotechnology, University of Gdansk, 80 822 Gdansk, Kladki 24, Poland
J Biol Chem 280:12292-8. 2005
..IbpA and IbpB form mixed complexes, and IbpA stimulates association of IbpB with substrate...
- Novel immobilized zinc(II) affinity chromatography for phosphopeptides and phosphorylated proteins
Department of Functional Molecular Science, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima 734 8551, Japan
J Sep Sci 28:155-62. 2005
..The affinity gel (Phos-tag agarose) was prepared by the quantitative reaction of N-hydroxysuccinimide-activated Sepharose and a Phos-tag derivative having a 2-aminoethylcarbamoyl group in dry CH3CN...
- Evaluation of a novel agarose-based synthetic ligand adsorbent for the recovery of antibodies from ovine serum
The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom
J Chromatogr B Analyt Technol Biomed Life Sci 860:209-17. 2007
..that the binding characteristics of the prototype adsorbent are similar to those obtained with Protein G Sepharose. This study indicates that the high-capacity prototype matrix is a feasible and potentially cost-effective ..
- Human serum IgM glycosylation: identification of glycoforms that can bind to mannan-binding lectin
James N Arnold
Medical Research Council Immunochemistry Unit, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom
J Biol Chem 280:29080-7. 2005
..Antigen-bound IgM pentamers therefore do not activate complement via the lectin pathway, but MBL might have a role in the clearance of aggregated IgM...
- Trypanosoma cruzi: partial characterization of minor cruzipain isoforms non-adsorbed to Concanavalin A-Sepharose
Vilma G Duschak
Instituto de Investigaciones Biotecnologicas, Universidad Nacional de General San Martin, Av Gral Paz y Albarellos, Parque Tecnológico Miguelete, INTI Edificio 24, 1650 San Martin, Prov Buenos Aires, Argentina
Exp Parasitol 104:122-30. 2003
..the partial characterization of a subset of atypical cruzipain molecules which do not bind to Concanavalin A-Sepharose column...
- Selective binding of the scavenger receptor C-type lectin to Lewisx trisaccharide and related glycan ligands
Peter J Coombs
Glycobiology Institute, Department of Biochemistry, University of Oxford, UK
J Biol Chem 280:22993-9. 2005
..These studies suggest that SRCL might be involved in selective clearance of specific desialylated glycoproteins from circulation and/or interaction of cells bearing Lewis(x)-type structures with the vascular endothelium...
- Isolation and characterization of a microsomal acid retinyl ester hydrolase
Phytonutrients Laboratory, Agricultural Research Service, United States Department of Agriculture, Beltsville, Maryland 20705, USA
J Biol Chem 280:23287-94. 2005
..Purification to homogeneity was achieved by sequential chromatography using SP-Sepharose cation exchange, phenyl-Sepharose hydrophobic interaction, concanavalin A-Sepharose affinity and Superose 12 gel ..
- An inter-subunit disulfide bond affects affinity of human lung extracellular superoxide dismutase to heparin
Department of Biochemistry, Hyogo College of Medicine, 1 1 Mukogawa cho, Nishinomiya, Hyogo 663 8501, Japan
Free Radic Res 37:823-7. 2003
..Moreover, the results obtained here also suggest that the redox state of the tissues might regulate the function of the EC-SOD...
- [Purification and identification of human recombinant IFN-beta expressed in yeast Pichia pastoris]
Qi wen Yu
Shanghai Institute of Immunology, Shanghai Second Medical University, Shanghai 200025, China
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 35:1035-9. 2003
..of purifying and identifying recombinant human IFN-beta (rhIFN-beta) expressed in yeast Pichia pastoris, Blue Sepharose 6 fast flow (Blue S6FF) and immunological affinity chromatography (IAC) were compared in this report...
- Purification of recombinant human carbonic anhydrase-II by metal affinity chromatography without incorporating histidine tags
Abir L Banerjee
Department of Chemistry and Molecular Biology, North Dakota State University, Fargo, ND 58105, USA
Protein Expr Purif 37:450-4. 2004
..We discovered that hCA-II preferentially binds to the Sepharose-iminodiacetate (IDA)-Zn(2+) column, and such binding does not require incorporation of either N- or C-terminal ..
- The preferred pathway of glycosaminoglycan-accelerated inactivation of thrombin by heparin cofactor II
Ingrid M Verhamme
Department of Pathology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 2561, USA
J Biol Chem 279:9785-95. 2004
..did not attenuate the DS-accelerated thrombin inactivation markedly, but DS displaced thrombin from heparin-Sepharose, suggesting that DS and heparin share a restricted binding site in or nearby exosite II, in addition to binding ..
- Sample displacement chromatography of Atlantic Salmon (Salmo salar) thrombin
Department of Chemistry, Biotechnology and Food Science, Agricultural University of Norway, Chr M Falsensgate 1, P O Box 5040, N 1432 As, Norway
J Biochem Biophys Methods 60:39-47. 2004
..chromatography system with non-homogenous sample application in the order of increasing affinity to Heparin Sepharose. The results suggest that this modified method could be useful in protein purification...
- Refolding and purification of recombinant human PDE7A expressed in Escherichia coli as inclusion bodies
Institute of Biochemistry, Medical Faculty, Institute of Biochemistry, University of Leipzig, Liebigstrasse 16, Leipzig, Germany
Protein Expr Purif 25:138-48. 2002
..5. The PDE7A1(147-482-His) construct could be purified after dialysis and concentration steps by either Zn2+-IDA-Sepharose chromatography or ResourceQ ion-exchange chromatography to homogeneity...
- Biochemical properties and cDNa cloning of two new lectins from the plasma of Tachypleus tridentatus: Tachypleus plasma lectin 1 and 2+
S C Chen
Institute of Biological Chemistry, Academia Sinica, Nankang Borough, Taipei 115, Taiwan, Republic of China
J Biol Chem 276:9631-9. 2001
A Sepharose CL-4B-binding protein, Tachypleus plasma lectin 1 (TPL-1), and a lipopolysaccharide (LPS)-binding protein, Tachypleus plasma lectin-2 (TPL-2), have been isolated from the plasma of Tachypleus tridentatus and biochemically ..
- Lactate dehydrogenase from the northern krill Meganyctiphanes norvegica: comparison with LDH from the Antarctic krill Euphausia superba
, , Poland
Comp Biochem Physiol B Biochem Mol Biol 128:233-45. 2001
..Antarctic krill isoenzyme exhibits high substrate affinities, high NAD inhibition, high inhibition at 10 mM pyruvate, lack of lactate inhibition, and high heat stability and resembles northern krill LDH-A*(4) isoenzyme...
- Phenylboronic acid-salicylhydroxamic acid bioconjugates. 2. Polyvalent immobilization of protein ligands for affinity chromatography
J P Wiley
Prolinx, Inc, 22322 20th Avenue SE, Bothell, Washington 98021, USA
Bioconjug Chem 12:240-50. 2001
..were compared with respect to the efficiency with which they were immobilized on salicylhydroxamic acid-modified Sepharose (SHA-X-Sepharose) by boronic acid complex formation. When immobilized on moderate capacity SHA-X-Sepharose (5...
- Novel lectin-like proteins on the surface of human monocytic leukemia cell line THP-1 cells that recognize oxidized cells
School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Japan
Arch Biochem Biophys 385:186-93. 2001
..macrophages were solubilized, and the membrane proteins obtained by affinity chromatographies using lactoferrin-Sepharose and band 3-Sepharose were purified by successive DE column chromatography and sodium dodecyl sulfate-..
- Comprehensive study on binding capacity of human immunoglobulin G to Avid AL affinity gel
J Y Shi
Unisyn Technologies, Inc, Tustin, CA 92680
J Chromatogr A 686:61-71. 1994
..2 mg/ml gel using binding buffer containing sodium sulfate. Recombinant Protein A gel was used as a control. In this study it also exhibited enhanced binding capacity with a high salt concentration binding buffer...
- Purification and characterization of the maize amyloplast stromal 112-kDa starch phosphorylase
H H Mu
Department of Food Science, Cook College, New Jersey Agricultural Experiment Station, Rutgers University, New Brunswick 08901, USA
Arch Biochem Biophys 388:155-64. 2001
..included ammonium sulfate fractionation, Sephacryl 300 HR chromatography, affinity starch adsorption, Q-Sepharose, and Mono Q chromatography...
- Protein adsorption on histidyl-aminohexyl-Sepharose 4B. II. Application to the negative one-step affinity purification of human beta2-microglobulin and immunoglobulin G
, , CNRS-UPRESA 6 022, France
J Chromatogr B Biomed Sci Appl 758:173-82. 2001
..from uremic patient's blood ultrafiltrate and plasma, respectively, was investigated on the histidyl-aminohexyl-Sepharose 4B adsorbent...
- Isolation of a novel agglutinin with complex carbohydrate binding specificity from fresh fruiting bodies of the edible mushroom Lyophyllum shimeiji
T B Ng
Department of Biochemistry, Chinese University of Hong Kong, Shatin, N T, Hong Kong, People s Republic of China
Biochem Biophys Res Commun 290:563-8. 2002
..The protein was adsorbed on CM-Sepharose even in 20 mM ammonium acetate (pH 5...
- Sephadex-based cell-affinity adsorbents: preparation and performance
Department of Transfusion Technology, CLB, Sanquin Blood Supply Foundation, P O Box 9190, 1066 AC Amsterdam, The Netherlands
Biotechnol Appl Biochem 35:55-60. 2002
..For comparison, cell binding to commercially obtained SpA-Sepharose was determined...