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



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Detection with Enzyme-Labeled Reagents

(Protocol summary only for purposes of this preview site)

This protocol describes chromogenic detection with alkaline phosphatase as well as both chromogenic and chemoluminescent methods of detection with horseradish peroxidase. Before the detection step, the nitrocellulose or PVDF membrane is incubated with the antigen using enzyme-conjugated primary antibodies or secondary detection reagents (such as species-specific anti-immunoglobulin antibodies, streptavidin, or Protein A) and washed. In this protocol, an appropriate substrate solution is applied to the membrane, and the signal is detected in the course of the enzymatic reaction, resulting either in development of insoluble colored products (chromogenic substrates) or in generation of light (chemiluminescent substrates).

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

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