[Histonet] Rat T Cell Marker

[Tarango, Mark]

Kristen & Ray,

I still say SATB1 is the way to go.  Kristen wants to stain T-cells that
don't express CD3 yet.  She'll need a T-cell marker that is expressed at
the earliest point possible (to include all T-cells, even CD3-negative
T-cells). 

Nearly every marker has some overlap with another lineage.  She probably
has already tried CD3, that being the most obvious and probably the
first thing anyone would think of, when trying to stain a T-cell (but it
ain't working out and she wants to know of other options).  I really
doubt she's working in a clinical rat lab.  She's doing research; but
even clinical labs end up using research antibodies.

I've used SATB1 on human and mouse and it works great once your protocol
is optimized.  Take it from someone who's actually used it.  

How would you tell the difference between something like ALK-negative
anaplastic large T-cell lymphoma and Hodgkin's lymphoma if the
morphology is similar and the tumor cells don't express mature B or
T-cell antigens?  You have to go for something that is expressed at an
earlier stage.  I'd chose PAX5 (B-cells) and SATB1 (T-cells) to be able
to figure it out.  CD3 is useless in such a situation.

This is research; don't be afraid to try something new to get you closer
to your answer.

Mark Adam Tarango HT(ASCP)
Histology & IHC Supervisor
Nevada Cancer Institute
One Breakthrough Way
Las Vegas, NV  89135
Direct Line (702) 822-5112
Treo (702) 759-9229
Fax (702) 939-7663
  

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
koellingr <@t> comcast.net
Sent: Wednesday, December 19, 2007 10:45 PM
To: Kristen Broomall; Histonet
Subject: Re: [Histonet] Rat T Cell Marker

Kristen,
To me something like CD3 is the way to go if I understand your project
as you described it.  Something like SATB1, which I've never used by IHC
but know something about, seems like overkill or using a good (but
incorrect) tool for the job.  SATB1 (special AT-rich binding protein 1)
I'm sure works well in Marks hands and others.  But it is a not well
characterized molecule being a loop-folding domain transcriptional
regulator and while true found "mainly" in thymocytes, there are
certainly indications of its presence in myeloid and erythroid
precursors as well as developing mouse CNS.  And it can be upregulated
and down-regulated.  And probably Marks statement that there might be
anti-rat SATB1 (and SATB2) antibodies around is true, but might not be
so easy to find or use.  Compare that to the absolutely, precisely
characterized and described gamma/delta/epsilon/zeta/eta CD3 TCR
complex.  It has been studied in minute detail all over the world, its
presence or absence in cell types 
is documented and redocumented countless times.  The number I've heard
is about 10,000 copies per T-cell and literally defines that cell.  If
you wanted to "mark" T-cells in flow, frozen or paraffin I think 99% of
labs would use CD3 at some point whether in periphery or thymus.  Well
characterized and protocoled anti-rat CD3 IHC antibodies exist in
multiple clones and from multiple vendors for rat IHC.  I like BD
Pharmingen but just my opinion. Without ever having stained for SATB1 in
rat, I believe what was said and guess yes you could do it, the question
is, is that the easiest and most efficient way to get to your answer?

Ray Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: "Kristen Broomall" <histotechkb <@t> gmail.com> 

> 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) 
> histotechkb <@t> gmail.com 
> _______________________________________________ 
> Histonet mailing list 
> Histonet <@t> lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
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"EMF <nvcancer.org>" made the following annotations.
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