Hi folks! I have come across an accident when I did my FACS intracellular staining: I completed the whole staining process but had to leave towards the end of the experiment, so I handed it over to my colleague. She washed the cells and resuspended them in PBS, but accidentally left the plate in the dark at room temperature overnight.

So my question is, can I still analyse these cells? Would it work if I redo the intracellular staining again and analyse? I used BD Cytofix (containing 4% paraformaldehyde) for fixation and homemade permeablisation buffer (0.5% Saponin, 1% BSA, 0.01% Sodium azide), and I stained the intracellular proteins first with unconjugated primary antibodies, then secondaries conjugated with AF647 or Dylight 650. For another group, I used primary antibodies directly conjugated with PE.

I would appreciate any suggestions! Thank you very much!