Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



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Cross-Linking Peptides to KLH with Maleimide

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KLH (keyhole limpet hemocyanin) is one of the most widely used carrier proteins (Harris and Markl 1999), which are complex molecules capable of eliciting a strong immune response in the host upon injection. Haptens, which are small antigens such as peptides and drug compounds, are very weakly or nonimmunogenic by themselves and require the assistance of a carrier protein to stimulate an immune response. KLHhapten conjugates are commonly used in antibody generation in a variety of hosts such as mice, rats, and rabbits. Because KLH is harvested from mollusks, it is physiologically distant from mammalian species and less likely to produce antibodies that cross-react mammalian antigens. Maleimide activation of carrier proteins makes it possible to conjugate sulfhydryl-containing haptens, and hence this chemistry is widely used for conjugating KLH with haptens (Lateef et al. 2007). Do not use Tris-based buffers because the primary amines will interfere with the conjugation reaction.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

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