Mattapallil jjmattapallil at UCDAVIS.EDU
Tue Sep 5 18:08:36 EST 2000

Pootr hi

I think it would be good idea to run PMA/Ionomycin stimulated samples
along side your actual samples as a positive control for your assay.
If you get a signal with stimulation at least you can be sure that
your staining protocol is working and what you see in unstimulated
samples maybe real. Also the differences between stimulated samples
from different treatments, if any, would indicate if the said
treatments were priming cells to produce certain cytokines.   Hope
that helps.

Joseph Mattapallil, B.V.Sc., Ph.D.

At 12:35 PM +0200 9/4/00, Piotr Wilczek wrote:
>I start my study of cytokine activation. The whole blood are derived
>from patient which undergo cardiac surgery operation with the us of
>Cardiac Pulmonary Bypass. The sample are taken : before anaesthesia,
>15 min of CPB, first and second postoperative day. I estimate the
>IL-4, IL-6, IL-8, MPO, TNF, IFN. I used the BD primary antibody for
>intracellular staining and DAKO IntraStain for fixation and
>permabilisation. All the sample are negative. But I'm not sure if
>the negative results are because there are really no activation of
>cytokine in this sample or because of wrong procedure. Should I
>pre-activate the lymphocyte (with BFA, Ionomycine) before the
>Many thanks for any suggestion
>Piotr Wilczek

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