John,

I don't shake and I don't dip... I blot.  According to what I was
taught, the blocking reagent does not adhere to anything in the tissues
and will be removed if rinsed. It needs the antibody or the core antigen
to form a complex with binding sites in the tissues.  BUT... I've always
wondered how that happens. If anyone can explain this, please do.  

Connie McManus


"John C. Dennis" wrote:
> 
> Dear Histofolk
> 
> Can anyone explain why now and again we hear/read that blocking solution
> should be shaken, not washed, from the tissue sections prior to
> applying primary or secondary solutions?
> 
> Once I was a shaker but there was no gift in simplicity.  I got uneven and
> patchy signal patterns.  Now I'm a dipper and no longer have those
> difficulties.
> 
> John Carroll Dennis
> Anatomy, Physiology, and Pharmacology
> 109 Greene Hall
> Auburn University, AL  36849

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