[Cytometry] PMA/Ionomycin Question

Beaumier, Coreen M CTR US USA MEDCOM WRAIR Coreen.M.Beaumier at us.army.mil
Fri May 6 09:05:12 EDT 2011


Dear all,

Lately, when using PMA/Ionomycin as a positive control for intracellular cytokine staining on human cells, we've been noticing something a bit strange. Sometimes our P/I stims worked and sometimes they didn't, depending on the donor.  We then tried two different concentrations (we've had success with both concentrations previously):

Concentration 1. 50ng/mL of PMA and 500ng/mL Ionomycin  or Concentration 2. 10ng/mL of PMA and 250ng/mL Ionomycin

What we found was that some donors like concentration 1 vs concentration 2 and others liked concentration 2 vs. concentration 1.  We also tried using new stocks of both our PMA and Ionomycin.  I'd like to avoid having to do both concentrations for each donor each time we run an experiment so does anyone have any suggestions?  Any advice would be greatly appreciated!


Thanks!
Coreen


Coreen M. Beaumier, PhD
Head, Immunology Section
Contractor, Clinical Research Management
Division of Viral Diseases
Walter Reed Army Institute of Research
Silver Spring, MD 20910
Phone: 301-319-9224
Fax: 301-319-9661





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