Re: CD4
From: | "C.M. vander Loos" <c.m.vanderloos@amc.uva.nl> |
> Following on from Nima's posting, I'll be staining cultured cells
>for CD1 and CD4 using Dako antibodies. Any suggestions for the preferred
>fixative would be welcome.
> Ian.
>
>
> Dr. Ian Montgomery,
> West Medical Building,
> University of Glasgow,
> Glasgow,
> G12 8QQ.
> Tel: 0141 339 8855. Extn:6602.
> Fax: 0141 330 2923
> e-mail: ian.montgomery@bio.gla.ac.uk
***********
Dear Ian,
Following our experiences with the immunostaining of intact cultured cells,
I think you should test both acetone (destroying the fatty membrane
structures) and 4% paraformaldehyde in PBS (2-5 min, RT). When using PFA
the addition of 0.1% saponin to reagents and washing buffers can be skipped
since this seems only useful for staining cytoplasmic antigens. Forget
about alcohol containing fixatives, as these will destroy the CD1/CD4
epitopes!
Good luck.
Chris
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