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We have used the J33 clone extensively in a variety of conjugates (FITC,
PE and PE:Cy5). It is an extremely reliable reagent and it is a
'recommended reagent' for use in the ISHAGE CD34+ cell enumeration
Guidelines.
Have you compared this reagent with any other pan-CD45 antibodies, such
as HLE-1 (BD) or KC56 (Coulter) to see if you detect similar events with
these clones?
Have you sorted these CD45- 'events' and had a look at the morphology of
them on the microscope?
How numerous are they and are they always present in different samples?
We do not usually see significant numbers of J33- events in our
analyses. Perhaps some of them are nucleated RBCs or megakaryocyte
fragments. Such particles and small platelet aggregates can exhibit the
light scatter characteristics of the events that you describe.
Do you have this problem in fresh samples only, or do you only find them
in apheresis concentrates that have been 'sitting around' for a while?
Hope this is helpful,
Rob Sutherland
ISHAGE Stem Cell Enumeration Committee,
Toronto Hospital
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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