Flow Cytometry on T Cells with IL-4, IFN-gamma- monensin use?!

1 post / 0 new
Gilligan
Gilligan's picture
Flow Cytometry on T Cells with IL-4, IFN-gamma- monensin use?!

I am totally new to research, i am an undergrad. I need some help!

I am trying to do intracellular staining of isolated T cells. I do not want to induce cytokine production with PMA/ionomycin however. I DO want to use monensin to prevent exocytosis of the cytokines already present!

How should I go about doing this? All the protocols I have found just say to add the monesin with the PMA, but i dont want to use the PMA.

ALSO, should i even be using monensin for IL-4 and IFN-gamma, or does brefeldin A work better?

I have not had success so far, while not using a protein transport blocker. I am hoping that the transport blocker is going to make a huge difference.

can anyone help? THANKS