Problem: diminished (barely detectable) calcium flux response to anti-CD3 in culture primary cells (mouse spleen) Other observations: indo-1 loading is a before; response to ionomycin is as expected. Instrument configuration, alignment, and laser power are all as before; anti-CD3 (Pharmingen) is new vial though is claimed to be same lot# and used at same concentration. History: cells from similar mice(same strain, age, isolation method) have given good responses in past; protocols for dye loading, cell incubation, antibody/agonist exposure, instrument setup are said by user to remain the same. Any thoughts? Dave Coder dcoder@u.washington.edu
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