IHC questions

From:"Kinsley, David" <david.kinsley@spcorp.com>

I'm currently trying to optimize an anti-BLC antibody on mouse pancreas
tissue.  In the lymph node I used as a positive control the signal was great
in the stromal cells of the follicle as expected.  (no background staining
at 5ug/ml) In my staining of pancreas tissue, the eyelets contain no
staining and there is very heavy background staining in the exocrine cells.
Does anyone have a suggestion as to blocking for this.  I'm using fresh
frozen tissue, cut at 10um, air dried 1 hour then cold acetone fixed for 20
minutes.  I block for peroxidase, avidin, biotin, and use a protein block
(10% serum of the second Ab species as well as Fetal calf serum, but no

Also, I'm considering an immunofluorescence stain on pancreas but would like
help as to reducing the background fluorescence found endogenously in the

Any suggestions are greatly appreciated.

Dave Kinsley MS,HT/HTL(ASCP) 

Schering-Plough Research Institute
Department of Immunology 
2015 Galloping Hill Road
Kenilworth, New Jersey 07033

Phone:  908-740-4669
Fax:      908-740-3084

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