Staining nuclear transcription factors for T helper subsets
Hello Forum,
I am curious whether anyone has been able to gate T helper subsets during phospho-CyTOF. Our protocol uses a 2% PFA fixation before surface staining, then methanol permeabilization before intracellular staining. Unfortunately, the surface markers CCR6 and CXCR3 are incompatible with fixation before staining. I was mainly wondering whether anyone has tried using any clones for the canonical T helper subsets (ie T-bet, Gata-3, and Ror-gt) to gate these subsets, how successful they were, or if there is a better solution.
Any help is very much appreciated,
Darius
I am curious whether anyone has been able to gate T helper subsets during phospho-CyTOF. Our protocol uses a 2% PFA fixation before surface staining, then methanol permeabilization before intracellular staining. Unfortunately, the surface markers CCR6 and CXCR3 are incompatible with fixation before staining. I was mainly wondering whether anyone has tried using any clones for the canonical T helper subsets (ie T-bet, Gata-3, and Ror-gt) to gate these subsets, how successful they were, or if there is a better solution.
Any help is very much appreciated,
Darius