what "pan T cell marker" are you using? Also, what do you define as an "immature" T cell? I've used BD Pharmingen mouse anti-rat CD3 in FFPE thymus with citrate retrieval and that will target the CD3 complex but a few other subsets of cells other than T-cells can express CD3. Also have used BD Pharmingen mouse anti-rat alpha/beta antibody on FFPE thymus with citrate retrieval but of course you will miss gamma/delta rearrangements but maybe that is not a concern. I've not tried the anti gamma/delta Ab on paraffin (only flow) but it might work. Perhaps others have tried. Also CD25 (low-affinity IL-2R) I've not had luck with on FFPE but maybe others have. Have had nice results with TdT (nuclear enzyme involved in TCR rearrangement) rabbit polyclonal on rat thymus FFPE. There are a few others, depends on the specificity and design of your model and exactly what you are looking for.
-------------- Original message --------------
From: "Kristen Broomall"
> Good Morning all, I was wondering if anyone had suggestions on an antibody
> to detect T cells (only, no B cells) in formalin fixed, paraffin embedded
> rat thymus and spleen. I have a Pan T Cell marker that is working well in
> the spleen, but so-so in the thymus, so I think that it is only staining the
> mature T cells, and not immature cells (which of course, is what we also
> need to see). I've been reading up alot on T cells and trying to figure this
> out (CD3, CD4, CD8, TCR, lions, tigers and bears - Oh My!), but I fear our
> deadline for getting staining done is shorter than the time I have to
> educate myself properly!
> Any suggestions would be greatly appreciated!
> Happy Holidays,
> Kristen Broomall, HT (ASCP)
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