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
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