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



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Biotinylating Antibodies Using Biotin Polyethylene Oxide (PEO) Iodoacetamide

(Protocol summary only for purposes of this preview site)

Biotinylation of free sulfhydryls can be effectively mediated using haloacetyl biotin derivatives (maleimide derivatives can also be used; see Protocol 4). To modify an antibody using this reagent, sulfhydryls must be available. Digestion of antibodies by the enzyme pepsin produces F(ab)2 fragments, which can be separated by mild reduction into two sulfhydryl-containing, univalent Fab fragments. Alternatively, thiol groups can be added by modifying amines with an appropriate cross-linker (see Protocol 6).

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

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