Yes,  I am following the procedure from the literature describing HIF1-alpha
staining in brain Tissue.
Cancer 2000 Jun 1;88(11):2606-18. Expression of hypoxia-inducible factor
1alpha in brain tumors: association with angiogenesis, invasion, and
progression. Zagzag D, Zhong H, Scalzitti JM, Laughner E, Simons JW, Semenza
GL
The antibody I am using is from Novus (catalog
# 100-123).  Staining requires HEIR,  which we perform in a microwave oven
(45 minutes 0.01M, pH6.0 citric acid buffer).  Antibody is incubated
overnight 4C (1:800). The monoclonal primary antibody is detected via
Tyramide
amplification (Dako CSA kit).  I know that these conditions are excessive,
and I have had some serious reservations about the validity of the staining
in other tissues. However,  some of our results have been correlated with
other markers that are associated with the expression of HIF-1.
Just last week I came across a reference (I have in PDF format) that
describes a procedure that is significantly less harsh.  I have not tried it
as of yet. If you like I could e-mail the reference and you could try that
procedure
Regards
Luis

--------------------------------------
Luis Chiriboga Ph.D., HT (ASCP) QIHC
New York University School Of Medicine
Kaplan Comprehensive Cancer Center
Bellevue Hospital
Department Of Pathology 4W27
27th Street & First Avenue
New York, N.Y. 10016

-----Original Message-----
From: Aneeta Patel [mailto:apatel@usuhs.mil]
Sent: Monday, July 09, 2001 12:47 PM
To: Histonet@pathology.swmed.edu
Subject: HIF


I am having no luck staining for hif alpha on paraffin embedded tissues
can anyone let me the details of the method,temp,incubation times so forth
and also a good positive control
thanks