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. 1973 Dec;135(4):867–873. doi: 10.1042/bj1350867

Polymerization of proteins with glutaraldehyde. Soluble molecular-weight markers

John W Payne 1,*
PMCID: PMC1165906  PMID: 4204954

Abstract

Glutaraldehyde is well known for its ability to react with proteins and to produce insoluble cross-linked aggregates. In contrast with this situation, conditions are described here which yield covalently linked soluble protein oligomers. The procedure is applicable to a wide range of proteins, and by slight variation in the reaction conditions, soluble polymers in the molecular weight range 3×104−2×107 were produced. The products are valuable aṡ molecular-weight markers, e.g. in sodium dodecyl sulphate–polyacrylamide-gel electrophoresis. The inherent similarities of these oligomers make them superior to commercial molecular-weight protein markers, which may have marked differences in composition and charge.

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Selected References

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  1. Andersson L. -O., Borg H., Mikaelsson M. Molecular weight estimations of proteins by electrophoresis in polyacrylamide gels of graded porosity. FEBS Lett. 1972 Feb 1;20(2):199–202. doi: 10.1016/0014-5793(72)80793-2. [DOI] [PubMed] [Google Scholar]
  2. Avrameas S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry. 1969 Jan;6(1):43–52. doi: 10.1016/0019-2791(69)90177-3. [DOI] [PubMed] [Google Scholar]
  3. Avrameas S., Taudou B., Chuilon S. Glutaraldehyde, cyanuric chloride and tetrazotized O-dianisidine as coupling reagents in the passive hemagglutination test. Immunochemistry. 1969 Jan;6(1):67–76. doi: 10.1016/0019-2791(69)90179-7. [DOI] [PubMed] [Google Scholar]
  4. Avrameas S., Ternynck T. The cross-linking of proteins with glutaraldehyde and its use for the preparation of immunoadsorbents. Immunochemistry. 1969 Jan;6(1):53–66. doi: 10.1016/0019-2791(69)90178-5. [DOI] [PubMed] [Google Scholar]
  5. Carpenter F. H., Harrington K. T. Intermolecular cross-linking of monomeric proteins and cross-linking of oligomeric proteins as a probe of quaternary structure. Application to leucine aminopeptidase (bovine lens). J Biol Chem. 1972 Sep 10;247(17):5580–5586. [PubMed] [Google Scholar]
  6. Dunker A. K., Rueckert R. R. Observations on molecular weight determinations on polyacrylamide gel. J Biol Chem. 1969 Sep 25;244(18):5074–5080. [PubMed] [Google Scholar]
  7. Fasold H., Klappenberger J., Meyer C., Remold H. Bifunctional reagents for the crosslinking of proteins. Angew Chem Int Ed Engl. 1971 Nov;10(11):795–801. doi: 10.1002/anie.197107951. [DOI] [PubMed] [Google Scholar]
  8. Griffith I. P. The effect of cross-links on the mobility of proteins in dodecyl sulphate-polyacrylamide gels. Biochem J. 1972 Feb;126(3):553–560. doi: 10.1042/bj1260553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Habeeb A. F. Preparation of enzymically active, water-insoluble derivatives of trypsin. Arch Biochem Biophys. 1967 Mar;119(1):264–268. doi: 10.1016/0003-9861(67)90453-5. [DOI] [PubMed] [Google Scholar]
  10. Habeeb A. J., Hiramoto R. Reaction of proteins with glutaraldehyde. Arch Biochem Biophys. 1968 Jul;126(1):16–26. doi: 10.1016/0003-9861(68)90554-7. [DOI] [PubMed] [Google Scholar]
  11. Hopwood D. A comparison of the crosslinking abilities of glutaraldehyde, formaldehyde and alpha-hydroxyadipaldehyde with bovine serum albumin and casein. Histochemie. 1969;17(2):151–161. doi: 10.1007/BF00277781. [DOI] [PubMed] [Google Scholar]
  12. Hopwood D., Callen C. R., McCabe M. The reactions between glutaraldehyde and various proteins. An investigation of their kinetics. Histochem J. 1970 Mar;2(2):137–150. doi: 10.1007/BF01003541. [DOI] [PubMed] [Google Scholar]
  13. Hopwood D. Fixatives and fixation: a review. Histochem J. 1969 May;1(4):323–360. doi: 10.1007/BF01003278. [DOI] [PubMed] [Google Scholar]
  14. Hopwood D. Some aspects of fixation with glutaraldehyde. A biochemical and histochemical comparison of the effects of formaldehyde and glutaraldehyde fixation on various enzymes and glycogen, with a note on penetration of glutaraldehyde into liver. J Anat. 1967 Jan;101(Pt 1):83–92. [PMC free article] [PubMed] [Google Scholar]
  15. Jansen E. F., Olson A. C. Properties and enzymatic activities of papain insolubilized with glutaraldehyde. Arch Biochem Biophys. 1969 Jan;129(1):221–227. doi: 10.1016/0003-9861(69)90169-6. [DOI] [PubMed] [Google Scholar]
  16. Ogata K., Ottesen M., Svendsen I. Preparation of water-insoluble, enzymatically active derivatives of subtilisin type Novo by cross-linking with glutaraldehyde. Biochim Biophys Acta. 1968 Jun 4;159(2):403–405. doi: 10.1016/0005-2744(68)90090-9. [DOI] [PubMed] [Google Scholar]
  17. Ottesen M., Svensson B. Modification of papain by treatment with glutaraldehyde under reducing and non-reducing conditions. C R Trav Lab Carlsberg. 1971;38(11):171–185. [PubMed] [Google Scholar]
  18. QUIOCHO F. A., RICHARDS F. M. INTERMOLECULAR CROSS LINKING OF A PROTEIN IN THE CRYSTALLINE STATE: CARBOXYPEPTIDASE-A. Proc Natl Acad Sci U S A. 1964 Sep;52:833–839. doi: 10.1073/pnas.52.3.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Richards F. M., Knowles J. R. Glutaraldehyde as a protein cross-linkage reagent. J Mol Biol. 1968 Oct 14;37(1):231–233. doi: 10.1016/0022-2836(68)90086-7. [DOI] [PubMed] [Google Scholar]
  20. Schejter A., Bar-Eli A. Preparation and properties of crosslinked water-insoluble catalase. Arch Biochem Biophys. 1970 Feb;136(2):325–330. doi: 10.1016/0003-9861(70)90202-x. [DOI] [PubMed] [Google Scholar]
  21. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  22. Wade H. E., Phillips B. P. Automated determination of bacterial asparaginase and glutaminase. Anal Biochem. 1971 Nov;44(1):189–199. doi: 10.1016/0003-2697(71)90360-5. [DOI] [PubMed] [Google Scholar]
  23. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  24. Weston P. D., Avrameas S. Proteins coupled to polyacrylamide beads using glutaraldehyde. Biochem Biophys Res Commun. 1971 Dec 17;45(6):1574–1580. doi: 10.1016/0006-291x(71)90200-2. [DOI] [PubMed] [Google Scholar]

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