Synopsis
Isoelectric focusing was used to determine the effects of fixation on the isoelectric point of bovine serum albumin. Aqueous solutions of formaldehyde, α-hydroxyadipaldehyde, osmium tetroxide and potassium dichromate produced no change or a relatively small decrease. Glutaraldehyde produced two fractions of fixed protein, the larger with a considerably lowered isoelectric point. The results are interpreted as suggesting that glutaraldehyde reacts rapidly and largely irreversibly, whereas the other fixatives react more slowly and reversibly with bovine serum albumin.
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References
Anderson, P. J. (1967). Purification and quantitation of glutaraldehyde and its effect on several enzyme activities in skeletal muscle.J. Histochem. Cytochem. 15, 652–61.
Baker, J. R. (1960).Principles of Biological Microtechniques. Methuen: London.
Bishop, D. H. L., Claybrook, J. R. &Spiegelman, S. (1967). Electrophoretic separation of viral nucleic acids on polyacrylamide gels.J. molec. Biol. 26, 373–87.
Bowes, J. H. &Cater, C. W. (1968). The interaction of aldehydes with collagen.Biochim. biophys. Acta 168, 341–52.
Bowes, J. H. &Kenten, R. H. (1949). The effect deamination and esterification on the reactivity of collagen.Biochem. J. 44, 142–52.
Filachione, E. M., Fein, M. L. &Harris, E. H. (1964). Tanning with glutaraldehyde.J. Am. Leath. Chem. Ass. 59, 281–92.
French, D. &Edsall, J. T. (1945). The reaction of formaldehyde with amino acids and proteins.Adv. prot. Chem. 2, 277–335.
Hake, T. (1965). Reactions of OsO4 and KMnO4 with amino acids, peptides and proteins.Lab. Invest. 14, 1208–12.
Hopwood, D. (1969a). A comparison of the crosslinking abilities of glutaraldehyde, formaldehyde and α-hydroxy-adipaldehyde with bovine serum albumin and casein.Histochemie 17, 151–61.
Hopwood, D. (1969b). Fixation of proteins by osmium tetroxide, potassium dichromate and potassium permanganate.Histochemie 18, 250–60.
Hopwood, D. (1970). The reactions between formaldehyde, glutaraldehyde and osmium tetroxide and their fixation effects on bovine serum albumin and on tissue blocks.Histochemie 24, 50–64.
Hopwood, D., Allen, C. R. &McCabe, M. (1970). The reactions between glutaraldehyde and various proteins. An investigation of their kinetics.Histochem. J. 2, 137–50.
Lenard, J. &Singer, S. J. (1968). Alterations of the conformation of proteins in red blood cell membranes and insolution by fixatives used in electron microscopy.J. Cell Biol. 37, 117–21.
Lillie, R. D. (1967). Hot chromation oxyphilia.Histochemie II, 332–49.
Pearse, A. G. E. (1968).Histochemistry: Theoretical and Applied, 3rd Ed., Vol. I. London: Churchill.
Porter, K. R. &Kallman, F. (1953). The properties and effects of osmium tetroxide as a tissue fixative with special reference to its use for electron microscopy.Expl Cell Res. 4, 127–41.
Quiocho, F. A. &Richards, F. M. (1964). Intermolecular crosslinking of a protein in the crystalline state: carboxypeptidase-A.Proc. natn. Acad. Sci. U.S.A. 52, 833–8.
Quiocho, F. A. &Richards, F. M. (1966). The enzymic behaviour of carboxypeptidase—A in the solid state.Biochemistry 5, 4062–76.
Schneider, G. &Schneider, G. (1967). Elektrophoretische Untersuchungen über die Wirkung von Gewebe-Fixierungsmitteln auf Eiweisslösungen.Histochemie 8, 132–44.
Seki, M. (1963). Zur physikalischen Chemie der histologischen Färbung IX. Über den Einfluss der Fixierung auf die Färbarkeit der histologischer Elemente.Z. Zellforsch. 18, 21–55.
Singer, M. &Morrison, P. R. (1948). The influence of pH, dye and salt concentration on the dye binding of modified and unmodified fibrin.J. biol. Chem. 175, 133–45.
Svensson, H. (1962). Isoelectric fractionation, analysis and characterization of ampholytes in natural pH gradients. II. Buffering capacity and conductance of isoionic ampholytes.
Tooze, J. (1964). Measurements of some cellular changes during the fixation of amphibian erthrocytes with osmium tetroxide solutions.J. Cell Biol. 22, 551–63.
Vesterberg, O. &Svennson, H. (1966). Isoelectric fractionation, analysis and characterization of ampholytes in natural pH gradients. Further studies on the resolving power in connection with separation of myoglobins.Acta Chem. Scand. 20, 820–34.
Young, E. G. (1963).Comprehensive Biochemistry (eds. M. Florkin & E. H. Stotz), Vol. 7, p. 25. Elsevier: New York.
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Hopwood, D. Use of isoelectric focusing to determine the isoeletric point of bovine serum albumin after treatment with various common fixatives. Histochem J 3, 201–205 (1971). https://doi.org/10.1007/BF01002564
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DOI: https://doi.org/10.1007/BF01002564