Hello everyone,
I am Doing Intracellular cytokine staining for IFN-g and IL-17. I want to stimulate cell (Isolated PBMC) with PMA, Ionomycine and Golgi Stop. After 5 h stimulation I want to block FC receptor. and then I will perform surface staining followed by Intracellular staining. But the problem is I can not able to do in one day. So, I decided to fix the cell with PFA. Can anyone please suggest me which step is the appropriate for fixation of the cell with PFA ? Is it a good idea to fix after adding FC block receptor?
If anyone please give me a suggestion regarding this issue, I will be highly benefited.
N.B. I am staining cell with CD3, CD4, CD8, CD14 & CD19
Best Regard
Razaul Haque