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I got out the AIS technical bulletin # 9001 and read the last paragraph in
which they discuss this phenomenon: After using the SBA- CD34 flasks,
"The ICH3 CD34 epitope is weakly detectable on the surface of the cells.
In addition, the CD34 antigen in weakly detectable with the monoclonal
antibodies MY10 and QBEND10. Other researchers have informed us that they
are better able to detect CD34 antigen on 70-90% of the purified cells by
using the antibodies 8G12 and 12.8: these antibodies are not commercially
available at this time." "See Technical Bulletin # 9002 for optimized
staining protocols." If you find a source for those other antibodies
mentioned above please let all of us know.
I have had the same trouble with my cells. One thing I have noticed is
that after about 7-10 days in culture the CD34 antigen becomes detectable
on an EPICS C again at low levels on a majority of the cells.
I just go on blind faith that I have some to start with after using the
cellector system and stimulate with growth factors appropriate for the
particular differentiation pathway I want. So far this has worked well. I
have had similar experiences with CD8+ and CD4+ cells, although they tend
to express new antigens more quickly than the stem cell.
The binding of antibody to antigens on cells may be disrupted by chemical
means or by slight changes in pH. These methods may affect the cells
adversely and should probably be avoided if the cells are to be expanded
after separation.
Hope this helps
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Roger A. Burger, Research Immunologist
Center for Persons with Disabilities
Utah State University, Logan, UT 84322-6800
Voice : (801)750-2042
Fax : (801)750-2044
InterNet SL061@cc.usu.edu
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