β-Galactosidase Labeling of Antibody Using MBS

  • G. Brian Wisdom
Part of the Springer Protocols Handbooks book series (SPH)


The heterobifunctional reagent, m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), is of value when one of the components of a conjugate has no free thiol groups, e.g., IgG. In this method (1), the IgG antibody is first modified by allowing the N-hydroxysuccinimide ester group of the MBS to react with amino groups in the IgG; the β-galactosidase is then added and the maleimide groups on the modified IgG react with thiol groups in the β-galactosidase to form thioether links. This procedure produces conjugates with molecular weights in the range 0.6–1 × 106.


Bovine Serum Albumin Sodium Phosphate Buffer Microbial Growth Thiol Group Free Thiol 
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  1. 1.
    O’Sullivan, M. J., Gnemmi, E., Morris, D., Chieregatti, G., Simmonds, A. D., Simmons, M., Bridges, J. W., and Marks, V. (1979) Comparison of two methods of preparing enzyme-antibody conjugates: application of these conjugates to enzyme immunoassay. Anal. Biochem. 100, 100–108.PubMedCrossRefGoogle Scholar
  2. 2.
    Carlson, J., Drevin, H., and Axen, R. (1978) Protein thiolation and reversible protein-protein conjugation, N-succinimidyl 3-(2-pyridyldithio)propionate—a new heterobifunctional reagent. Biochem. J. 173, 723–737.Google Scholar
  3. 3.
    Pierce Catalog and Handbook (1994) Pierce, Rockford, IL.Google Scholar
  4. 4.
    Ellman, G. L. (1959) Tissue sulfhydryl groups. Arch. Biochem. Biophys. 82, 7–75.CrossRefGoogle Scholar
  5. 5.
    Kastenschmidt, L. L., Kastenschmidt, J., and Helmrich, E. (1968) Subunit interactions and their relationship to the allosteric properties of rabbit skeletal muscle phosphorylase b. Biochemistry 7, 3590.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • G. Brian Wisdom
    • 1
  1. 1.Division of Biochemistry, School of Biology and BiochemistryThe Queen’s University, Medical Biology CentreBelfastUK

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