[Cytometry] PMA/Ionomycin Question

Apostolos Bossios apostolos.bossios at lungall.gu.se
Sat May 7 11:35:56 EDT 2011


Dear Coreen,

The theory back PMA/Ion stimulation is that it will increase the expression of genes that have already have been stimulated. That is why is used in a lot of studies as a stimulus.
We use the first dose. So I think that the fact that some donors are not stimulated in natural

Best
Apostolos

************************** 
Apostolos Bossios MD, PhD 

University of Gothenburg 
Krefting Research Centre
Box 424
SE-405 30  Gothenburg 
Sweden 
 
Visiting address: Medicinaregatan 1G 
Tel: +46 (0) 31-786 6719
Fax: +46 (0) 31-786 6730
e-mail: apostolos.bossios at gu.se




-----Original Message-----
From: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Beaumier, Coreen M CTR US USA MEDCOM WRAIR
Sent: den 6 maj 2011 15:05
To: Cytometry at lists.purdue.edu
Subject: [Cytometry] PMA/Ionomycin Question

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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