Measurement of T and B cell turnover with bromodeoxyuridine.
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.
Tough
David F
DF
The Scripps Research Institute, La Jolla, California.
Sprent
Jonathan
J
The Scripps Research Institute, La Jolla, California.
Stephens
Geoffrey L
GL
NIAID, Bethesda, Maryland.
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D001402:B-Lymphocytes / Q000166:cytology*; D001973:Bromodeoxyuridine; D002453:Cell Cycle; D002469:Cell Separation; D003584:Cytological Techniques; D004261:DNA Replication; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D007158:Immunologic Techniques; D051379:Mice; D051381:Rats; D013194:Staining and Labeling; D016176:T-Lymphocyte Subsets / Q000166:cytology; D013601:T-Lymphocytes / Q000166:cytology*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D005456:Fluorescent Dyes; D001973:Bromodeoxyuridine