Flow labelling for Common Gamma chain of IL-2 receptor

From: Belinda Kramer (BelindaK@chw.edu.au)
Date: Wed Nov 28 2001 - 00:58:14 EST


Dear all,

I am trying to label human peripheral blood lymphocytes to detect expression
of the common gamma chain of the IL-2, IL-4..etc receptor. I normally just
lyse out red cells from whole blood and wash white cells with PDS / Azide
(0.1%)/ BSA (0.1%) before labelling.

 I'm using a rat-anti human monolconal-Biotin and then Streptavidin-Fitc,
and am having problems with consistent labelling.  That is, sometimes the
lymphocytes label, othertimes not.  I've tried various temperatures (RT, 4C,
on ice), and tried titering the antibody over a wide range to optimise
labelling, but haven't been able to come up with a consistently good result.

Does anyone have any experience with trying to label this antigen?  Or is it
more likely to be  a problem with the antibody?

Many thanks

Belinda Kramer
Oncology Research Unit
The Children's Hospital at Westmead
Australia
Belindak@chw.edu.au


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