CTL assay

Priti Kumar priti at mcbl.iisc.ernet.in
Thu Mar 15 08:28:21 EST 2001


Hi,
	I am having a lot of troubl loading up chromium into B-
lymphoblastoid cells for use in a cytotoxic T cell assay. I am using 4 to
5 million cells in 200ul of RPMI-10 (human AB) with HEPES in the presence
of 0.25mCi of chromium at 37 deg. celsius for 1 hour. Under these
conditions the amount of label in 1000 viable cells after lysis with 1%
Triton X 100 is barely 1500 counts. The spontaeneous release after 4 hours
of incubation is also about the  same. Can someone suggest a method of
getting optimal incorporation?

PRITI KUMAR
DEPARTMENT OF MICROBIOLOGY AND CELL BIOLOGY
INDIAN INSTITUTE OF SCIENCE
BANGALORE
INDIA-560012.
PHONE NO. (080)3092685
FAX NO. 91-80-3602696
e-mail: priti at mcbl.iisc.ernet.in



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