UNIT 7.13 Measurement of Polyclonal Immunoglobulin Synthesis Using the Reverse Plaque Technique

  1. Giovanna Tosato1,
  2. Sandra E. Pike1,
  3. Michael Blaese2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im0713s00

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Tosato, G., Pike, S. E. and Blaese, M. 2001. Measurement of Polyclonal Immunoglobulin Synthesis Using the Reverse Plaque Technique. Current Protocols in Immunology. 00:II:7.13:7.13.1–7.13.6.

Author Information

  1. 1

    Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland

  2. 2

    National Cancer Institute, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: DEC 1991


This unit describes the reverse hemolytic plaque assay, an effective method for measuring the number of immunoglobulin (Ig)-secreting cells present in a cell population at any particular time. Cell populations that can be assayed using the technique include peripheral blood mononuclear cells or cells from tissues such as the tonsils. The basic protocol is divided into three stages. First, protein A-sensitized sheep red blood cells (SRBC), guinea pig complement, and anti-Ig antibody are prepared. Test samples are then combined and incubated with the SRBC, complement, and antibody in appropriate chambers. Finally, the resulting plaques are scored. A support protocol describes the preparation of plaquing chambers.