[Histonet] placental IHC for the novice

[Patsy Ruegg]

Julienne,
DAKO will provide you with a free handbook on IHC methods.
Patsy

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Julienne
Rutherford
Sent: Thursday, October 07, 2004 2:48 PM
To: Histonet <@t> Lists. Utsouthwestern. Edu
Subject: [Histonet] placental IHC for the novice 


Hello. I'm new to histonet and am not trained in histo or IHC, but am
very interested. I'm throwing myself into the fray, and hope you can
help me. In response, which I welcome privately or in the forum, please
use simple language for I am but a simple woman with a simple dream of
studying monkey placentas.  I want to study IGF-1 localization in the
marmoset monkey placenta. I'm doing a lot of reading regarding methods,
but I need help as to what I need to do (and by extension, how to ask
for the money to do it in grant applications!). The folks in our histo
lab are going to help me and train me but I want to come to them more
prepared and I have to provide the specific materials I'll need. I have
both 10% NBF-fixed tissues (most sitting around in formalin for a very
long time), and I also have frozen, but by this I mean stuck in the
regular old lab freezer to freeze conventionally. I will begin
snap-freezing placentae we collect from now on, but what can I get out
of this old stuff? Can I thaw out what I have and snap-freeze them in
OCT? Should I try to do IHC on both the formalin-fixed AND the frozen
tissue?

In order to build a budget I need to know everything I need to make
these slides. So, what kind of slides and coverslips do I need, what are
all the chemicals involved, what is a secondary antibody and why do I
need it, is a chromagen different from a stain, etc.? Also, is there
something akin to and "IHC for Dummies" resource? I've looked at
ihcworld.com for protocols but it's over my head at this point. I
recognize that this is a skill perfected over many years, but I really
would like a primer on the basics so I can have a more informed
discussion about it.

If the elementary pitch of my questions is inappropriate to the forum, I
sincerely apologize. I'm just not sure where else to turn. Please
respond to my Texas e-mail if you'd prefer to school (or scold) me
privately!

Thanks in advance,
Julienne Rutherford

Research Assistant (Adjunct)
Southwest Foundation for Biomedical Research
Southwest National Primate Research Center
P.O. Box 760549
San Antonio, Texas 78245-0549
phone (210) 258-9864
fax (210) 258-9883
jrutherford <@t> icarus.sfbr.org

Ph.D. candidate
Indiana University
Bloomington, Indiana
jnruther <@t> indiana.edu _______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet