Unit

UNIT 4.7 Measurement of T and B Cell Turnover with Bromodeoxyuridine

  1. David F. Tough1,
  2. Jonathan Sprent1,
  3. Geoffrey L. Stephens2

Published Online: 1 MAY 2007

DOI: 10.1002/0471142735.im0407s77

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Tough, D. F., Sprent, J. and Stephens, G. L. 2007. Measurement of T and B Cell Turnover with Bromodeoxyuridine. Current Protocols in Immunology. 77:4.7:4.7.1–4.7.9.

Author Information

  1. 1

    The Scripps Research Institute, La Jolla, California

  2. 2

    NIAID, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2007
  2. Published Print: MAY 2007

Abstract

This unit describes two methods used to determine the rate of turnover of T and B cells in vivo. In each method, this is accomplished by administering 5-bromo-2′-deoxyuridine (BrdU), a thymidine analog, and measuring the rate at which cells become labeled with this DNA precursor during the S phase of the cell cycle. Cells that contain BrdU are then detected in vitro using fluorochrome-labelled anti-BrdU monoclonal antibody by flow cytometry. The unit includes an Alternate Protocol that may be useful in situations where the Basic Protocol proves incompatible with particular antibody-fluorochrome conjugates or other downstream staining procedures. Using this procedure, a wide range of antibody-fluorochrome combinations can be used.