Unit

UNIT 5.6 Production of Antipeptide Antisera

  1. John E. Coligan1,
  2. James P. Tam2,
  3. Jun Shao (MAP systems)2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142301.ns0506s00

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Coligan, J. E., Tam, J. P. and Shao, J. 2001. Production of Antipeptide Antisera. Current Protocols in Neuroscience. 00:5.6:5.6.1–5.6.21.

Author Information

  1. 1

    National Institute of Allergy and Infectious Diseases, Bethesda, Maryland

  2. 2

    Vanderbilt University, Nashville, Tennessee

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: SEP 1997

Abstract

This unit describes methods used for the chemical coupling of synthetic peptides to carrier proteins, required for the preparation of peptide immunogens. The carrier protein described here is keyhole limpet hemocyanin (KLH) because it is the one most commonly used. However, other proteins may be used in place of KLH, including bovine serum albumin (BSA) and ovalbumin. Coupling may be accomplished as described with MBS, which requires a Cys residue in the peptide, or with glutaraldehyde, EDCI, or BDB. Also included are an assay for detecting free sulfhydryl (SH) groups, a means of calculating coupling efficiency, an immunization schedule, an indirect ELISA, and a method for preparing a peptide affinity column. The final methodology described is the multiple antigen (MAP) system.