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We too have found that it is not possible to label the CD34+ selected cells
with anti-CD34, and we have not been able to convincingly detect mouse Ig on
the
cells either (I'm pretty sure it is a mouse monoclonal). We have tried holding
the cells at 37 degrees to let them re-express CD34
with only fair results (the usual problems with cell death and high
background).
We have convinced ourselves that we are indeed getting mostly CD34+ cells
because:
1) They form CFU's
2) They have the right light scatter pattern
3) They are markedly enriched for CD34 message by RT-PCR, and the non-adherent
cells
do not have detectable CD34 message.
I too would be interested in a good way to re-label the CD34+ cells - it would
be a heck of lot
easier than RT-PCR!!
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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