Ca flux in mice protocol

shawn_jackson shawn_jackson at student.hms.harvard.edu
Wed Mar 20 16:07:14 EST 2002


hello all -

I'd like to do some Ca flux experiments in mouse splenocytes.
It looks like loading with indo-1 is the way to go, and stimulating with
anti-CD3e clone 145-2C11.

Does anyone have a protocol they're willing to share?
In the literature I've seen indo-1 between 1 and 5 uM, and anti-CD3
between 1 and 90 ug/ml.

I'm hoping to do direct ex vivo studies on CD8 cells. We're thinking to
stimulate them to flux by addition of MHC I tetramer and sort by 62L
expression (using 4 color flow).

Can anyone give advice as to what anti-CD8 or anti-CD62L clones WON'T
stimulate Ca flux?

And finally, has anybody tried this sort of thing on PBMCs?


Thanks a bunch

Shawn Jackson
Div. Viral Pathogenesis
Beth Israel Deaconess Medical Center
Boston, MA 02115
617-667-0182





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