Immunohistochemistry Problem


Hi all,

I'm a student currently doing a research project in IHC with GADD45 and Cdc2
on cell pellets with known antigen.

The antibody I'm using is anti-GADD45 (H-165), SC-797, Lot I187 rabbit polyclonal
IgG from Santa Cruz and anti-Cdk1/Cdc2 lot C12720 from Transduction Laboratorires.
 The detection kit is from Biogenex - the StrAviGen Multilink Kit.  I use a
30 minute heat antigen retrieval in 0.1M citrate buffer (microwave).  The antibody
concentration has been varied at 1/500, 1/100, and 1/50 with 24hour incubations
at 4 degrees.  No results have been observed, no background, just blue cells
(hematoxylin).  All reagents have been checked previously and work with other
antibodies from a routine lab (University of Alberta Hospital), and procedures
were exactly the same.  Has anyone worked with this antibody or encountered
this situation?  What was done to solve the problem?

Any suggestions would be appreciated!


<< Previous Message | Next Message >>