agarose chromatography


Summary: A method of gel filtration chromatography using agarose, the non-ionic component of agar, for the separation of compounds with molecular weights up to several million.

Top Publications

  1. Rodriguez P, Ruiz M, Price G, Zannis Hadjopoulos M. NAP-2 is part of multi-protein complexes in HeLa cells. J Cell Biochem. 2004;93:398-408 pubmed
    ..This fraction, although able to supercoil relaxed DNA, did so to a lesser extent than the Q-Sepharose fraction. The data suggest that NAP-2 is in complex(es) with other proteins, which are distinct from histones. ..
  2. Onoue S, Nemoto Y, Harada S, Yajima T, Kashimoto K. Human antithrombin III-derived heparin-binding peptide, a novel heparin antagonist. Life Sci. 2003;73:2793-806 pubmed
    ..The results provided new findings about the structure-activity relationship of hAT III (123-139), and led us to the successful development of a potent antagonist for heparin. ..
  3. Kavanagh Wright L, Smith T, Gibney J, McKenna T. Characterization of macroprolactin and assessment of markers of autoimmunity in macroprolactinaemic patients. Clin Endocrinol (Oxf). 2009;70:599-605 pubmed publisher
    ..Comprehensive analysis of macroprolactin confirmed its composition as an IgG molecule or fragment with a PRL molecule. The occurrence of macroprolactin does not appear to be associated with autoimmunity. ..
  4. Hsu F, Ho T, Lai C, Lin C, Chen H. Cloning, sequencing, expression, and single-step purification of the adenosylcobinamide kinase/adenosylcobinamide-phosphate guanylyltransferase (CobU) from Salmonella typhimurium ATCC 19585. Protein Expr Purif. 2005;42:178-81 pubmed
    ..typhimurium ATCC 19585 and, without introducing a purification tag sequence to the N- or C-terminus of the recombinant enzyme, a new single-step purification method based on hydrophobic interaction chromatography. ..
  5. Clifton J, Huang F, Gaso Sokac D, Brilliant K, Hixson D, Josic D. Use of proteomics for validation of the isolation process of clotting factor IX from human plasma. J Proteomics. 2010;73:678-88 pubmed publisher
    ..Our methodology is very helpful for further process optimization, rapid identification of target proteins with relatively low abundance, and for the design of subsequent steps for their removal or purification. ..
  6. Wade P, Shaffer S, Jaehning J. Resolution of transcription factors from a transcriptionally active whole-cell extract from yeast: purification of TFIIB, TBP, and RNA polymerase IIa. Protein Expr Purif. 1993;4:290-7 pubmed
    ..In addition, the three previously described isoforms of yeast RNA polymerase II were resolved and form IIa, the intact, unphosphorylated isoform proposed to be involved in initiation, was purified to apparent homogeneity. ..
  7. Finn R, Kapelioukh I, Paine M. Rainbow tags: a visual tag system for recombinant protein expression and purification. Biotechniques. 2005;38:387-8, 390-2 pubmed
    ..This technology, which we call Rainbow Tagging, facilitates expression and downstream processing of recombinant proteins, paving the way for the development of automated high-throughput protein expression systems. ..
  8. Watanabe R, Samusawa Saito R, Oshima Y. Development of saxitoxin-conjugated affinity gels. Bioconjug Chem. 2006;17:459-65 pubmed
    ..Finally, the STX-conjugated affinity gel was effective for concentrating STX-binding proteins from pufferfish and bullfrog plasma. ..
  9. Talhouk R, Maa ni F, Kalaa ni N, Zoubian G, Simaa n C, Abi Sai d M, et al. Partial purification and characterization of proteins with growth promoting activities from ovine mammary gland secretions. Domest Anim Endocrinol. 2001;21:143-59 pubmed
    ..The factors modulate mammary cell function and may play a role in mammary gland development. ..

More Information


  1. Biswas E. Nucleotide binding domain 1 of the human retinal ABC transporter functions as a general ribonucleotidase. Biochemistry. 2001;40:8181-7 pubmed
    ..Thus, the nucleotide binding preference of NBD1 is as follows: CTP > GTP > ATP >> UTP. These studies demonstrate that NBD1 of ABCR is a general nucleotidase, whereas NBD2 is a specific ATPase...
  2. Borisova A, El darov M, Zhgun A, Aleksandrova S, Omel ianiuk N, Sokov B, et al. [Purification and properties of recombinant Erwinia carotovora L-asparaginase expressed in E.coli cells]. Biomed Khim. 2003;49:502-7 pubmed
    ..Physical-chemical and structural properties of recombinant Erwinia carotovora L-asparaginase are similar to the enzymes from the wild strains Erwinia carotovora and recombinant L-asparaginase Erwinia chrysanthemi. ..
  3. Siu S, Boushaba R, Topoyassakul V, Graham A, Choudhury S, Moss G, et al. Visualising fouling of a chromatographic matrix using confocal scanning laser microscopy. Biotechnol Bioeng. 2006;95:714-23 pubmed
    ..The competitive adsorption of BacLight Red-labelled whole cells or cell debris and PicoGreen-conjugated dsDNA was also visualised using CSLM. ..
  4. Tiainen P, Gustavsson P, Ljunglöf A, Larsson P. Superporous agarose anion exchangers for plasmid isolation. J Chromatogr A. 2007;1138:84-94 pubmed
    ..Two strategies were evaluated, one with and one without enzymatic digestion of RNA. The strategy without RNase gave high plasmid recovery, quantitative removal of protein and a 70% reduction in RNA. ..
  5. Iijima N, Uchiyama S, Fujikawa Y, Esaka M. Purification, characterization, and molecular cloning of group I phospholipases A2 from the gills of the red sea bream, Pagrus major. Lipids. 2000;35:1359-70 pubmed
    ..These results demonstrate that three group I PLA2, G-1, G-2, and G-3 PLA2, are expressed in the gill filaments of red sea bream. ..
  6. Mortensen K, Hansen H, Grentzmann G, Buckingham R, Sperling Petersen H. Osmo-expression and fast two-step purification of Escherichia coli translation termination factor RF-3. Eur J Biochem. 1995;234:732-6 pubmed
    ..All assays showed that the purified RF-3 was highly active with a specific activity of approximately 2000 units/mg. ..
  7. Wanderley M, Udrisar D, Martins M, Antunes Rodrigues J. Propranolol stimulates histone phosphorylation by a putative PK-C in partially purified homogenate of rat testicular interstitial cells. A possible mechanism for increased testosterone secretion by propranolol. Horm Metab Res. 2000;32:259-64 pubmed
    ..The ability of propranolol to activate the putative protein kinase C could be related to its stimulatory effect on testosterone secretion by Leydig cells. ..
  8. He W, Bruley D. Soft gel chromatography column analysis and design for the production of high molecular weight blood factors: model molecule Protein C. Adv Exp Med Biol. 2003;510:121-5 pubmed
  9. Paludan Müller C, Gram L, Rattray F. Purification and characterisation of an extracellular fructan beta-fructosidase from a Lactobacillus pentosus strain isolated from fermented fish. Syst Appl Microbiol. 2002;25:13-20 pubmed
    ..The enzyme had temperature and pH optima at 25 degrees C and 5.5, respectively. At concentrations of up to 12% NaCl no adverse effect on the enzyme activity was observed. ..
  10. Berna P, Berna N, Porath J, Oscarsson S. Comparison of the protein adsorption selectivity of salt-promoted agarose-based adsorbents. Hydrophobic, thiophilic and electron donor-acceptor adsorbents. J Chromatogr A. 1998;800:151-9 pubmed
  11. Tomoyasu A, Higashio K, Kanomata K, Goto M, Kodaira K, Serizawa H, et al. Platelet-rich plasma stimulates osteoblastic differentiation in the presence of BMPs. Biochem Biophys Res Commun. 2007;361:62-7 pubmed
    ..These results suggested that platelets contain not only growth factors for proliferation but also novel potentiator(s) for BMP-dependent osteoblastic differentiation. ..
  12. Yamamoto Y, Takahashi I, Ogata N, Nakazawa K. Purification and characterization of N-acetylglucosaminyl sulfotransferase from chick corneas. Arch Biochem Biophys. 2001;392:87-92 pubmed
    ..It appears that this enzyme competes with beta-galactosyltransferase in binding to the nonreducing GlcNAc residue on KS synthesis; this suggests that the sulfation of the GlcNAc residue is coupled with the elongation of the KS chain. ..
  13. Ogino H, Hiroshima S, Hirose S, Yasuda M, Ishimi K, Ishikawa H. Cloning, expression and characterization of a lipase gene (lip3) from Pseudomonas aeruginosa LST-03. Mol Genet Genomics. 2004;271:189-96 pubmed
    ..However, in the presence of most of the organic solvents tested in this study (the exceptions were ethylene glycol and glycerol), the stability of the Lip3 lipase was lower than that of the LST-03 lipase. ..
  14. Wu J, Eyre D. Matrilin-3 forms disulfide-linked oligomers with matrilin-1 in bovine epiphyseal cartilage. J Biol Chem. 1998;273:17433-8 pubmed
    ..The results indicate a molecular stoichiometry of (matrilin-1)2(matrilin-3)2. ..
  15. Kiefer P, Mathieu M, Close M, Peters G, Dickson C. FGF3 from Xenopus laevis. EMBO J. 1993;12:4159-68 pubmed
    ..Increasing the contribution from mouse FGF3 led to a more restricted host range for the chimeric ligand. ..
  16. Herrero Turrion M, Rodriguez R, Velasco A, Gonzalez Sarmiento R, Aijon J, Lara J. Growth hormone expression in ontogenic development in gilthead sea bream. Cell Tissue Res. 2003;313:81-91 pubmed
    ..These results suggest that the transcription of GH is involved in the early developmental stages of sea bream and the asynchronous growth-related changes are not due to distinct distribution of GH cells. ..
  17. Sigdel T, Cilliers R, Gursahaney P, Crowder M. Fractionation of soluble proteins in Escherichia coli using DEAE-, SP-, and phenyl sepharose chromatographies. J Biomol Tech. 2004;15:199-207 pubmed
    ..This work describes an easy, inexpensive way to fractionate the soluble proteins in E. coli and a way to better profile the E. coli proteome. ..
  18. Strongin A, Gorodetsky D, Stepanov V. The study of Escherichia coli proteases. Intracellular serine protease of E. coli-an analogue of bacillus proteases. J Gen Microbiol. 1979;110:443-51 pubmed
    ..coli protease was similar to that of the Bacillus licheniformis enzyme. These data indicate a relationship between proteolytic enzymes of evolutionary distant Gram-negative Enterobacteriaceae and Gram-positive spore-forming Bacillus. ..
  19. Bhattacharya A, Arya R, Clark C, Ackers J. Absence of lipophosphoglycan-like glycoconjugates in Entamoeba dispar. Parasitology. 2000;120 ( Pt 1):31-5 pubmed
    ..No LPG-like molecules could be identified in E. dispar in contrast to E. histolytica, suggesting that these molecules may be absent in the non-pathogenic species. ..
  20. Pérez A, Rucavado A, Sanz L, Calvete J, Gutierrez J. Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper. Braz J Med Biol Res. 2008;41:12-7 pubmed
    ..e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the ;gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms. ..
  21. Mora J, Garrido A, Antonijuan A, Martinez S, González Sastre F. Applicability of alpha-fetoprotein-concanavalin A (AFP-ConA) binding to discriminate between germinal or hepatic origin of AFP in germ cell tumour patients during chemotherapy or follow-up. Clin Chem Lab Med. 2007;45:932-3 pubmed
  22. Brandt P, Vanaman T. Studies of calmodulin-dependent regulation. Methods Mol Biol. 2004;284:111-27 pubmed
    ..The assay of CaM Kinase II using a synthetic peptide substrate is also described. ..
  23. Achouri Y, Noel G, Vertommen D, Rider M, Veiga da Cunha M, van Schaftingen E. Identification of a dehydrogenase acting on D-2-hydroxyglutarate. Biochem J. 2004;381:35-42 pubmed
    ..The corresponding human protein was expressed in HEK-293 cells and it was shown to catalyse the detritiation of DL-2-hydroxy[2-3H]glutarate with similar properties as the purified rat enzyme. ..
  24. McCardell B, Kothary M, Hall R, Sathyamoorthy V. Identification of a CHO cell-elongating factor produced by Vibrio cholerae O1. Microb Pathog. 2000;29:1-8 pubmed
    ..Partly purified Cef did not increase cyclic AMP or prostaglandin E(2)levels in CHO cells which suggests that its mechanism of action differs from that of cholera toxin. ..
  25. Lubick K, Burgess D. Purification and analysis of a phospholipase A2-like lytic factor of Trichomonas vaginalis. Infect Immun. 2004;72:1284-90 pubmed
    ..These results suggest that LF is a PLA(2) and may be an important virulence factor of T. vaginalis mediating the destruction of host cells and contributing to tissue damage and inflammation in trichomoniasis. ..
  26. Sridevi N, Srivastava S, Khan B, Prabhune A. Characterization of the smallest dimeric bile salt hydrolase from a thermophile Brevibacillus sp. Extremophiles. 2009;13:363-70 pubmed publisher
    ..To our knowledge, this is the first report describing the purification of BSH to homogeneity from a thermophilic source. ..
  27. Fosang A, Last K, Maciewicz R. Aggrecan is degraded by matrix metalloproteinases in human arthritis. Evidence that matrix metalloproteinase and aggrecanase activities can be independent. J Clin Invest. 1996;98:2292-9 pubmed
    ..ITEGE374 neoepitope. The results suggest that in some circumstances, despite extensive proteolysis of the core protein, aggrecan molecules may be cleaved by MMPs or aggrecanase in the interglobular domain, but not both. ..
  28. Böckmann R, Dickneite C, Goebel W, Bohne J. PrfA mediates specific binding of RNA polymerase of Listeria monocytogenes to PrfA-dependent virulence gene promoters resulting in a transcriptionally active complex. Mol Microbiol. 2000;36:487-97 pubmed
    ..By using EMSA and DNaseI footprint experiments it could be shown that PrfA leads to an enhanced and specific binding of RNAP. Transcriptional activity of RNAP in vitro, using the actA promoter, was strictly dependent on PrfA. ..
  29. Murakami K, Haneda M, Qiao S, Naruse M, Yoshino M. Prooxidant action of rosmarinic acid: transition metal-dependent generation of reactive oxygen species. Toxicol In Vitro. 2007;21:613-7 pubmed
    ..Cytotoxicity of rosmarinic acid may be related to the prooxidant action resulting from metal-reducing activity. ..
  30. Sheng Z, Chang S, Chirico W. Expression and purification of a biologically active basic fibroblast growth factor fusion protein. Protein Expr Purif. 2003;27:267-71 pubmed
    ..The ability of the purified GST-bFGF to stimulate the growth of human umbilical vein endothelial cells (HUVECs) was equivalent to that of purified recombinant 18 kDa bFGF. ..
  31. Doonan S, Cutler P. General strategies. Methods Mol Biol. 2004;244:1-13 pubmed
  32. Edelstein C, Italia J, Klezovitch O, Scanu A. Functional and metabolic differences between elastase-generated fragments of human lipoprotein[a] and apolipoprotein[a]. J Lipid Res. 1996;37:1786-801 pubmed
    ..The comparable size of apo[a] fragments observed in the urine of humans and injected mice invites the speculation that enzymes of the elastase family may play a role in the biology of Lp[a] in vivo. ..
  33. Valladares L, Erices A, Lioi X, Iturriaga H. Characterization of the oligosaccharides of plasma sex hormone binding globulin from noncirrhotic alcoholic patients. Steroids. 2000;65:275-80 pubmed
    ..communis-I indicated that at least the SHBG in this fraction, from alcoholics or controls, contained two glycosylation sites and that the sites were differentially glycosylated. ..
  34. Ono M, Okuda K, Takazoe I. Purification and characterization of a thiol-protease from Bacteroides gingivalis strain 381. Oral Microbiol Immunol. 1987;2:77-81 pubmed
    ..The isoelectric point was 4.9. The enzyme activity was enhanced by dithiothreitol, L-cysteine, and 2-mercaptoethanol and inhibited by p-chloromercuribenzoic acid, N-ethylmaleimide, and iodoacetic acid. ..
  35. Ding X, Li C, Sun L, Wang H. [Expression, purification and identification of human matrix metalloproteinase-2]. Sheng Wu Gong Cheng Xue Bao. 2001;17:643-7 pubmed
    ..The recombinant MMP-2 with 50 kD molecular weight is smaller than natural MMP-2, which suggested degradation occurred to it. ..
  36. Laurence A, Astoul E, Hanrahan S, Totty N, Cantrell D. Identification of pro-interleukin 16 as a novel target of MAP kinases in activated T lymphocytes. Eur J Immunol. 2004;34:587-97 pubmed
    ..The present study thus reveals how phosphoproteomic analysis opens previously unrecognized avenues for research, and yields novel insights about targets for MAP kinases in T lymphocytes. ..
  37. Mascher E, Lundahl P. Sodium dodecyl sulphate-protein complexes. Changes in size or shape below the critical micelle concentration, as monitored by high-performance agarose gel chromatography. J Chromatogr. 1989;476:147-58 pubmed
    ..The formation of all five SDS-protein complexes investigated was practically completed at cmcSDS. ..
  38. Yonezawa Y, Tokunaga H, Ishibashi M, Tokunaga M. Characterization of nucleoside diphosphate kinase from moderately halophilic eubacteria. Biosci Biotechnol Biochem. 2001;65:2343-6 pubmed
    ..This protein showed anomalous mobility on SDS-PAGE which is typical of a halophilic protein. It was stable and active over a wide range of salt concentrations, from 0 to 4.0 M NaCl. ..
  39. Pearlstone J, Smillie L. Skeletal muscle troponin C. Expression and purification of the recombinant intact protein and its isolated N- and C-domain fragments. Methods Mol Biol. 2002;172:161-74 pubmed
  40. Valente F, Saraiva L, LeGall J, Xavier A, Teixeira M, Pereira I. A membrane-bound cytochrome c3: a type II cytochrome c3 from Desulfovibrio vulgaris Hildenborough. Chembiochem. 2001;2:895-905 pubmed
    ..With the periplasmic [Fe] hydrogenase TpII-c3 is reduced much slower than TpI-c3, and no catalytic effect of TpI-c3 is observed. ..
  41. Baluja Conde I, Brito Moreno A, Acosta Bas C, Zalueta Rodríguez O, Melchor Rodríguez A, Amores Sanchéz I, et al. Monoclonal antibody against free beta-subunit of human chorionic gonadotropin. Hybrid Hybridomics. 2002;21:381-4 pubmed
    ..The affinity constant of this MAb was 1.5 x 10(10)M(-1). ..
  42. Antonov Y, Dmitrochenko A, Leontiev A. Interactions and compatibility of 11 S globulin from Vicia faba seeds and sodium salt of carboxymethylcellulose in an aqueous medium. Int J Biol Macromol. 2006;38:18-24 pubmed
  43. Lucena S, Guerrero B, Salazar A, Gil A, Arocha Piñango C. Degradation of extracellular matrix proteins (fibronectin, vitronectin and laminin) by serine-proteinases isolated from Lonomia achelous caterpillar hemolymph. Blood Coagul Fibrinolysis. 2006;17:427-35 pubmed
    ..achelous caterpillar. ..
  44. Urbatsch I, Wilke Mounts S, Gimi K, Senior A. Purification and characterization of N-glycosylation mutant mouse and human P-glycoproteins expressed in Pichia pastoris cells. Arch Biochem Biophys. 2001;388:171-7 pubmed
    ..Mass spectrometry indicated that mutant and wild-type proteins did not differ significantly in mass, demonstrating that the wild-type proteins contain no N-glycosylation. ..
  45. Meyer B, Duvillard L, Owen A, Packard C, Caslake M. Fractionation of cholesteryl ester rich intermediate density lipoprotein subpopulations by chondroitin sulphate. Atherosclerosis. 2007;195:e28-34 pubmed
    ..It remains to be determined if these cholesterol rich IDL particles are potentially more atherogenic than the triglyceride rich IDL particles. ..
  46. Chou K, Kukhanova M, Cheng Y. A novel action of human apurinic/apyrimidinic endonuclease: excision of L-configuration deoxyribonucleoside analogs from the 3' termini of DNA. J Biol Chem. 2000;275:31009-15 pubmed
    ..This discovery suggests that APE1 could be critical for the activity of l-OddC or other l-nucleoside analogs and may play additional important roles in cells that were not previously known. ..
  47. Hall R, Embery G, Waddington R. Modification of the proteoglycans of rat incisor dentin-predentin during in vivo fluorosis. Eur J Oral Sci. 1996;104:285-91 pubmed
  48. Nunes C, Dias Cabral A. Angiotensin I retention behavior on Butyl-Sepharose under linear loading chromatographic conditions. J Chromatogr A. 2009;1216:2332-8 pubmed publisher
    ..The retention of the cis form of angiotensin I increases with the increase in salt concentration in the mobile phase and seems to be little affected by temperature. ..
  49. Nóvoa Valiñas M, Perez Lopez M, Melgar M. Comparative study of the purification and characterization of the cytosolic glutathione S-transferases from two salmonid species: Atlantic salmon (Salmo salar) and brown trout (Salmo trutta). Comp Biochem Physiol C Toxicol Pharmacol. 2002;131:207-13 pubmed
  50. Copeland P, Driscoll D. Purification and analysis of selenocysteine insertion sequence-binding protein 2. Methods Enzymol. 2002;347:40-9 pubmed
  51. Shende N, Upadhye V, Kumar S, Gangane N, Harinath B. Study of M. tuberculosis ES-31 and ES-20 antigen levels in different pathogenic grades of lymph node tuberculosis. Int J Tuberc Lung Dis. 2007;11:222-6 pubmed
    ..05). Elevated levels of immune complexed ES-20 antigen in patient's serum may be a useful immunological marker for weak immune response patients in lymph node TB. ..
  52. Zanette D, Soffientini A, Sottani C, Sarubbi E. Evaluation of phenylboronate agarose for industrial-scale purification of erythropoietin from mammalian cell cultures. J Biotechnol. 2003;101:275-87 pubmed
    ..By adding an anion exchange chromatography step and gel filtration for polishing, a pure and active product can easily be obtained by an integrated, start-to-end process optimized for industrial-scale operations. ..
  53. Kim K, Lee B, Pou S, Ciccolella D. Effects of activation of polymorphonuclear leukocytes on airway goblet cell mucin release in a co-culture system. Inflamm Res. 2003;52:258-62 pubmed
    ..In the co-culture system, activation of PMN results in release and degradation of mucins, both of which are almost entirely accounted for by serine proteases but not other cellular products such as superoxide. ..