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foxp3 staining

PostPosted: Mon Aug 07, 2017 5:21 pm
by Kjwaller

I have done a bit of cytof already and have been fairly happy with it but now I'm changing project. I need to do my surface stain, phospho stain and also foxp3 stain. Previously I stimulated, add 1% PFA, surface stain, methanol and then phospho stains, this worked fine but I'm now unsure how to do my foxp3 staining. I know the methanol messes with the surface stain which is why I did this order but I have also found the stat1 signal is quite reduced when I use a foxp3 perm buffer. Any suggestions for the best method? I'm assuming methanol won't be enough for foxp3 as it's nuclear?

Thanks in advance,

Re: foxp3 staining

PostPosted: Thu Sep 28, 2017 3:50 pm
by SamuelKD
Hi Kate,

I can tell you from data that I've been looking at the foxp3 antibody sold by fluidigm on 162Dy, clone PCH101 ( ... y--50tests) appears fine on methanol staining. There is some question about background non-specific binding, but I'm sure others would be able to address that concern - the samples I've seen were sorted for CD4+. Mike from the HIMC once told me that they saw high background on monocytes, but that wasn't a concern for the researcher they were working with. Other clones apparently had staining on all cell types including B cells.

Hope this helps,