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Alkaline Phosphatase Labeling of Antibody Using Glutaraldehyde

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

Abstract

The chemistry of the homobifunctional reagent, glutaraldehyde, is complex. It reacts with the amino and, to a lesser extent, the thiol groups of proteins, and when two proteins are mixed in its presence, stable conjugates are produced without the formation of Schiff bases. Self-coupling can be a problem, unless the proteins are at appropriate concentrations. In the method (1) described, there is usually little self-coupling of the enzyme or the IgG antibody. However, the size of the conjugate is large (>106 Daltons), since several molecules of each component are linked. This is the simplest labeling procedure to carry out and, although the yields of enzyme activity and immunoreactivity are small, the conjugates obtained are stable and practical reagents.

Keywords

Schiff Base Ammonium Sulfate Thiol Group Short Glass Free Antibody 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Engvall, E. and Perlmann, P. (1972) Enzyme-linked immunosorbent assay, ELISA. III. Quantitation of specific antibodies by enzyme-labelled anti-immunoglobulin in antigen-coated tubes. J. Immunol. 109, 129–135.PubMedGoogle Scholar
  2. 2.
    Avrameas, S. and Ternynck, T. (1971) Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penetration. Immunochemistry 8, 1175–1179.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 Biological CentreBelfastUK

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