Problems identifying RANTES and TGF-β1 population with the FlowCytomix Pro

[Katherine Garro]

Hello,

I´ve used the Human Rantes Simplex BMS8287FF Kit and the Human TGF-β1 BMS8249FF Kit in addition to the Human Th1/Th2 kit in the same assay. I don´t have the Rantes and TGF-β1 manual , only the Th1/Th2 manual, for the population setup to choose the correct one at the A and B populations.

Now that I´m trying to make the standard curve using the FlowCytomix Pro 3.0, I can`t identified the RANTES and TGF-β1 populations from the others included in the other kit. Anybody has the manual? or could give some advice.

Besides this problem, when I run the samples for the Human Th1/Th2 Kit almost all the cytokines values are in red. Have anybody had an idea of what can I do to correct this, I already have eliminated some of the standard values on the curve so my Fit value reach at leat 90% but I keep having the same problem.

In attached an image of my A and B populations. At B population I have two population at the same range of Fluorescent Intensity and i can`t separated them.

Hope someone could help me

Thanks!