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foxp3 staining

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Kjwaller

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Posts: 11

Joined: Sat Aug 27, 2016 6:56 am

Post Mon Aug 07, 2017 5:21 pm

foxp3 staining

Hi,

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,
Kate
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SamuelKD

Participant

Posts: 4

Joined: Tue Dec 15, 2015 4:42 pm

Post Thu Sep 28, 2017 3:50 pm

Re: foxp3 staining

Hi Kate,

I can tell you from data that I've been looking at the foxp3 antibody sold by fluidigm on 162Dy, clone PCH101 (https://www.fluidigm.com/reagents/prote ... 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,

Sam

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