Abstract
The most popular methods for estimation of molecular weights of proteins are gel filtration (GF) and polyacrylamide gel electrophoresis (PAGE) (see Appendix for a glossary of terms used in this chapter). These two approaches may employ aqueous media or denaturing solvents such as solutions of urea, guanidine, sodium dodecylsulfate (SDS), sodium decylsulfate (NaDS), or other detergents. The usual practice is to construct a “calibration curve” and to utilize linear interpolation for estimation of Stokes’ radius, molecular weight, or log molecular weight. This chapter provides a review of the various types of calibration curves available, with their theoretical and empirical bases, and attempts to provide a comprehensive and unified approach to analysis of data from gel filtration, PAGE, and SDS-PAGE.
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References
Ackers, G. K. (1964). Biochemistry 3:723–730.
Ackers, G. K. (1967). J. Biol. Chem. 242:3237–3238.
Andrews, P. (1965). Biochem. J. 96:595–606.
Banker, G. A., and Cotman, C. W. (1972). J. Biol. Chem. 247:5856–5861.
Cantz, M., Chrambach, A., and Neufeld, E. F. (1970). Biochem. Biophys. Res. Commun. 39:936.
Chrambach, A., and Rodbard, D. (1971). Science 172:440–451.
Corvol, P., Rodbard, D., Bardin, C. W., and Chrambach, A. (1971). J. Biol. Chem. 246:3435–3443.
Davis, B. K. (1974). Proc. Natl. Acad. Sci. USA 71: 3120–3123.
Determann, H. (1968). Gel Chromatography, Gel Filtration, Gel Permeation, Molecular Sieves: A Laboratory Handbook, Springer-Verlag, New York, 195 pp.
Fawcett, J. S., and Morris, C. J. O. R. (1966). Sep. Sci. 1:9.
Ferguson, K. A. (1964). Metabolism 13:985–1002.
Finney, D. J. (1964). Statistical Method in Biological Assay, 2nd ed., Griffin, Cambridge, 668 pp.
Finney, D. (1971). Probit Analysis, Cambridge University Press, Cambridge.
Fish, W. W., Reynolds, J. A., and Tanford, C. (1970). J. Biol. Chem. 245:5166–5168.
Fisher, L. (1969). In Laboratory Techniques in Biochemistry and Molecular Biology, Vol. I (T. S. Work and Work, E., eds.), North-Holland, Amsterdam, p. 157.
Fisher, M. P., and Dingman, C. W. (1971). Biochemistry 10:1895–1899.
Frank, R. N., and Rodbard, D. (1975). Arch. Biochem. Biophys. 171:1–13.
Gonnene, A., and Lebowitz, J. (1975). Anal. Biochem. 64:414–424.
Hedrick, J. L., and Smith, A. J. (1968). Arch. Biochem. Biophys. 126:155–164.
Henckes, G., Crochet, M., Labedaw, B., and Legault-Demare, J. (1974). Anal. Biochem. 60:1–14.
Kawasaki, T., and Ashwell, G. (1976). J. Biol. Chem. 251:1296–1302.
Laurent, T. C., and Killander, J. (1964). J. Chromatogr. 14:317–330.
Maizel, J. V. (1971). In Methods in Virology, Vol. 5 (K. Maramorosch and H. Koprowski, eds.), Academic Press, New York, 179–246.
Martin, R. G., and Ames, B. N. (1961). J. Biol. Chem. 236:1372.
Morawetz, H. (1966). Macromolecules in Solution, Interscience New York, p. 359.
Morris, C. J. O. R. (1966). Protides Biol. Fluids 14:543–551.
Morris, C. J. O. R., and Morris, P. (1971). Biochem. J. 124:517–528.
Neville, D. M. (1971). J. Biol. Chem. 246:6328–6334.
Ogston, A. G. (1958). Trans. Faraday Soc. 54:1754–1757.
Ogston, A. G. (1966). Br. Med. Bull. 22:105–108.
Ogston, A. G. (1970). J. Phys. Chem. 74:668–669.
Ogston, A. G., and Phelps, C. F. (1961). Biochem. J. 78:827–833.
Ornstein, L. (1964). Ann. N.Y. Acad. Sci. 121:321–349.
Ostrowski, W., and Wasyl, Z. (1969). Biochim. Biophys. Acta 181:479–482.
Porath, J. (1963). Pure Appl. Chem. 6:233–246.
Reddy, N. B., Engel, W. K., and Festoff, B. W. (1976). Biochim. Biophys. Acta (in press).
Reynolds, J. A., and Tanford, C. (1970a). J. Biol. Chem. 244:5161–5165.
Reynolds, J. A., and Tanford, C. (1970b). Proc. Natl. Acad. Sci. USA 66:1002.
Richards, E. G., and Lecanidou, R. (1974). In Electrophoresis and Isoelectric Focusing in Polyacrylamide Gel (R. C. Allen and H. R. Maurer, eds.), De Gruyter, New York, pp. 16–22.
Rodbard, D., and Chrambach, A. (1970). Proc. Natl. Acad. Sci. USA 65:970–977.
Rodbard, D., and Chrambach, A. (1971). Anal. Biochem. 40:95–134.
Rodbard, D., and Chrambach, A. (1974). In Electrophoresis and Isoelectric Focusing in Polyacrylamide Gel (R. C. Allen and H. R. Maurer, eds.), De Gruyter, New York, pp. 28–62.
Rodbard, D., and Hutt, D. M. (1974). In Proceedings, Symposium on Radioimmunoassay and Related Procedures in Medicine, International Atomic Energy Agency, Vienna, pp. 165–192.
Rodbard, D., Kapadia, G., and Chrambach, A. (1971). Anal. Biochem. 40:135–157.
Rodbard, D., Levitov, C., and Chrambach, A. (1972). Sep. Sci. 7:705–723.
Segrest, J. P., and Jackson, R. L. (1972). Meth. Enzymol. 28:54–63.
Shapiro, A. L., and Maizel, J. V. (1969). Anal. Biochem. 29:505–514.
Shapiro, A. L., Vinuela, E., and Maizel, J. V. (1967). Biochem. Biophys. Res. Commun. 28:815–820.
Siegel, L. M., and Monty, K. J. (1966). Biochim. Biophys. Acta 112:346–362.
Sober, H. A. (1970). Handbook of Biochemistry, Chemical Rubber Co., Cleveland.
Squire, P. G. (1964). Arch. Biochem. 107:471–478.
Tanford, C. (1961). Physical Chemistry of Macromolecules, Wiley, New York.
Tanford, C., Nozaki, Y., Reynolds, J. A., and Makino, S. (1974). Biochemistry 13:2369–2376.
Ugel, A. R., Chrambach, A., and Rodbard, D. (1971). Anal. Biochem. 43:410–426.
Whitaker, J. R. (1963). Anal. Chem. 35:1950–1952.
White, M. L., and Dorion, G. H. (1961). J. Polymer Sci. 55:731–740.
Wold, S. (1974). Technometrics 16:1–11.
Zwaan, J. (1967). Anal. Biochem. 21:155–168.
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Rodbard, D. (1976). Estimation of Molecular Weight by Gel Filtration and Gel Electrophoresis. In: Catsimpoolas, N. (eds) Methods of Protein Separation. Biological Separations. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-9984-1_3
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DOI: https://doi.org/10.1007/978-1-4684-9984-1_3
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