[Histonet] Re: which one first - antigen retrieval or endo peroxidase block?

From:Brian Chelack



Ahh but it can matter!!

 

The antigen retrieved epitope (enzymatic retrieval in this instance)
recognized by the clone 15C5 for Bovine Viral Diarrhea (BVD) is destroyed by
methanol. Thus if you are using hydrogen peroxide in methanol to block your
endogenous peroxidases after performing antigen retrieval on a positive BVD
case you will obtain a very weak or even negative result. It appears that
the formalin cross linking protects the epitope from the effects of the
methanol. So run your assay both ways if you are concerned, trust your
observations.

 

I learned this the hard way back in 1989.

 

  If you would be a real
seeker after truth, it is necessary that at least once in your life you
doubt, as far as possible, all things. 

Rene Descartes   (1596
- 1650)

 

Brian Chelack
Special Projects Manager
Prairie Diagnostic Services Inc.
52 Campus Drive, Saskatoon, SK S7N 5B4
Phone (306) 966-7211
Fax (306) 966-7244
Email brian.chelack@pds.usask.ca 

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