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I have a colleague who wants to pulse lab animals with BrdU, then do
successive blood draws and follow appearance of BrdU in various subsets of
PBMC's. He wants to label the blood cells with anti-BrdU:FITC and cell
surface antigens with PE (CD4:PE, etc.). He has been able to demonstrate
BrdU incorporation into cultured cell lines, but has failed to get positive
results with fresh PBMC's. Also, cell surface antigen staining has not
worked as well in his ethanol-fixed (following surface antigen staining;
pre-BrdU staining) samples as in PBMC's stained only for surface markers
and then paraformaldehyde fixed.
Has anyone successfully stained PBMC's for BrdU and surface markers at the
same time? We would appreciate any protocols or tips that you might
suggest.
Thanks in advance.
* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
* Richard K. Meister Email: Meister.1@osu.edu *
* The Ohio State University Voice: (614) 292-9716 *
* Dept. of Veterinary Biosciences FAX: (614) 292-6473 *
* Cytometry Instrumentation Lab *
* 1925 Coffey Road *
* Columbus, OH 43210 U.S.A. *
* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web