Summary
The crosslinking effects of formaldehyde, α-hydroxyadipaldehyde and glutaraldehyde have been compared by various techniques. Using a micro-Ouchterlony technique with an aldehyde treated bovine serum albumin-rabbit anti-bovine serum albumin system it was found that glutaraldehyde prevented precipitin line formation except at very high titres of antibody. The effects of formaldehyde and α-hydroxyadipaldehyde were less marked. Cellulose acetate electrophoresis of aldehyde treated bovine serum albumin showed an increase in mobility compared with the untreated protein. Starch gel electrophoresis of aldehyde fixed liver slices showed no protein loss after glutaraldehyde fixation whereas the other aldehydes permitted proteins to be extracted. Polyacrylamide gel electrophoresis of the aldehyde treated bovine serum albumin showed a little change in mobility after formaldehyde and α-hydroxyadipaldehyde treatment and a little polymer formation. Glutaraldehyde on the other hand produced much polymer. These findings were confirmed by gel filtration with Sephadex G-200. Intermolecular crosslinking with glutaraldehyde was dependant on the aldehyde concentration.
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References
Andrews, P.: Estimation of the molecular weights of proteins by Sephadex Gel-filtration. Biochem. J. 91, 222–233 (1964).
—: Molecular sieve chromatography applied to molecular size and molecular weight estimation. Lab. Pract. 16, 851–856 (1967).
Boedtker, H.: The reaction of ribonucleic acid with formaldehyde. 1. Optical absorbance studies. Biochemistry 6, 2718–2728 (1967).
Bowes, J. A., and C. W. Cater: Crosslinking of collagen. J. appl. Chem. (Lond.) 15, 296–304 (1965).
Davis, B. J.: Disc electrophoresis. II. Method and application to human serum proteins. Ann. N.Y. Acad. Sci. 121, 404–427 (1964).
Davis, P., and B. E. Tabor: Kinetic study of the crosslinking of gelatin by formaldehyde and glyoxal. J. polymer Sci. A 1, 799–815 (1963).
Eränkö, O., A. Kokko, and U. Söderholm: Histochemical evidence of three types of esterases in the adrenal medulla of the rat. Histochemie 2, 383–388 (1962).
Flodin, P.: Dextran gels and their applications in gel filtration. Dissertation, 85 pp. Uppsala: AB Pharmacia 1962.
Gustavson, K. H.: The chemistry of tanning processes. New York: Academic Press 1956
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. (Lond.) 101, 83–92 (1967a).
—: The behaviour of various glutaraldehydes on Sephadex G 10 and some implications for fixation. Histochemie 11, 289–295 (1967b).
Hopwood , D., and M. McCabe: The reaction between glutaraldehyde and various proteins. An investigation of their kinetics. In preparation (1968).
Janigan, D. T.: Tissue enzyme fixation studies. 1. The effects of aldehyde fixation on β-glucuronidase, β-galacrosidase, N-Acetyl-B-glucosaminidase and β-glucosidase in tissue blocks. Lab. Invest. 13, 1038–1050 (1964).
—: The effect of aldehyde fixation on acid phosphatase activity in tissue blocks. J. Histochem. 13, 473–483 (1965).
Maunsbach, A. B.: The influence of different fixatives and fixation methods on the ultrastructure of rat kidney proximal tubule cells. 11. Effects of varying osmolarity ionic strength, buffer system and fixative concentration of glutaraldehyde solutions. J. Ultrastruct. Res. 15, 283–309 (1966).
Milgrom, F., and E. Witebsky: Immunological studies on adrenal gland. 1. Immunization with adrenals of foreign species. Immunology 5, 46–66 (1962).
Ornstein, L.: Disc electrophoresis. 1. Background and theory. Ann. N.Y. Acad. Sci. 121, 321–349 (1964).
Pearse, A. G. E.: Histochemistry, theoretical and applied, 3rd ed., vol. 1. London: Churchill 1968.
Peters, T., and C. A. Ashley: An artefact in radioautography due to binding of free amino acids to tissues by fixatives. J. Cell Biol. 83, 53–60 (1967).
Quiocho, F. A., and F. M. Richards: Intermolecular crosslinking of a protein in the crystalline state: carboxypeptidase-A. Proc. nat. Acad. Sci. (Wash.) 52, 833–839 (1964).
—: The enzymic behaviour of carboxypeptidase-A in the solid state. Biochemistry 5, 4062–4076 (1966).
Sabatini, D. D., K. Bensch, and R. J. Barnett: Cytochemistry and electron microscopy. The preservation cellular ultrastructure and enzymatic activity by aldehyde fixation. J. Cell Biol. 17, 19–58 (1963).
Smith, I.: Chromatographic and electrophoretic techniques. London: Heinemann 1960.
Smithies, O.: Zone electrophoresis in starch gels and its application to studies of serum proteins. Advanc. Protein Chem. 14, 65–113 (1959).
Tooze, J.: Measurements of some cellular changes during the fixation of amphibian erythrocytes with osmium tetroxide solutions. J. Cell Biol. 22, 551–563 (1964).
Tristram, G. R., and R. H. Smith: The amino acid composition of some purified proteins. Advanc. Protein Chem. 18, 277–318 (1963).
Wold, F.: Bifunctional reagents. In: Methods in enzymology, vol. XI, 617–640, ed. by C. H. W. Hirs. London: Academic Press 1967.
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Hopwood, D. A comparison of the crosslinking abilities of glutaraldehyde, formaldehyde and α-hydroxyadipaldehyde with bovine serum albumin and casein. Histochemie 17, 151–161 (1969). https://doi.org/10.1007/BF00277781
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DOI: https://doi.org/10.1007/BF00277781