Dear Rumi,

First of all I would like to clarify that if you want to perform intracellular staining for IL-4 or not? If yes, then I would like to say that measuring IL-4 intracellularly is a limitation in flow cytometry. You cant get the proper result. This problem persists for a long time. But in serum or plasma, bead array can detect IL-4. Anyway if you stimulate the cells with recombinant IL-4 or free fatty acid followed by short boost with PMA + ionomycin, this can help. If you more information, please tell, I will let you know as much as i can.

Thanks,

Debanjan

 Thank you for your kind and prompt resp.

I am very interested in your experience of IL-4 and intracellular staining!

I'd like also if you agree to share with you some preliminar data.

I wonder if there is any other mediator by PMA and ionomycin that could help.

Gabriele

Dear Rumi,

OK, I have no problem if you want to share some preliminary data of yours with me. I have worked on human leishmaniasis and found that intracellular IL-4 is undetectable by FACS calibur cytometer from BD with their Intracellular cytokine kit. Then we contacted Perdue university who are experts of flow. They told us that this is indeed a major limitation of flow cytometry that of IL-4 detection intracellularly.

My email id is [email protected]. If you want to share or ask anything you can write to me also.

Thanks