Help! CD3+ Flow Cytometry

SNOWTREK snowtrek at aol.com
Sat Aug 10 16:02:48 EST 1996


"human peripheral Blood lymphocytes has a very bright and tight CD3
staining pattern...I'm not too surprised with your results on cell lines,
have you tried peripheral blood T cells?"......We normally use the Jurkat
E6-1 line (from ATCC) and as negative control Jurkat J.RT3. Another line
which is positive for CD3 is MOLT16. The lines should be free of
mycoplasma and recloned (high CD3 producers). CD3 is unstably expressed in
many cell lines...."

First 2 responses: Yeah, but my purpose is a standardized, quantitatively
well-characterized assessment of this anti-CD3 mab's binding. I need to be
able to walk over to the incubator and pull out a T-175  full of cell XYZ,
grab a quarter million, mix in the mab, and get reasonable reproducibility
in response. The variability and poor characterization of PBLs (possible
pathogen present?, etc) makes them a non-viable alternative. Third
response: The Jurkats didn't work for me. As you suggest, recloning them
might allow isolation of a CD3 dense, homogeneous poulation. And, I'll
look into the Molt16s. THANKS for the replies!!!!



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