Re: Primary antibody incubation

From:Victoria Baker

Fernando,
My answer to this is it depends, I don't believe there
really is a universal "best".  I have worked in
clinical, industrial and currently in academia, I
handle human and animal tissue/cells and do a large
variety of IHC/IF/ISH/FISH work.  I will tell you now
that primary incubation times are never written in
stone because they are only ONE part of the procedure.
 Each lab has to work out its own conditions based on
the variables they have to work with.  Both the
conditions and variables differ from lab to lab.

In my lab currently we run PCNA, bcl-2, Cyclin D1,
GSTpi, 3-clones of p53, and BrdU at one to two hours
incubation at RT.
All NFkB subunits, 8oxodG, BPDE, Cox 1 & 2, p21,
3-nitrotyrosine and MGMT overnight at 4 C.

Just name a few.

Vikki Baker
Institute for Cancer Prevention
Valhalla, New York 10595


--- Fernando Capela e Silva  wrote:
> What is the best in immnuohistochemestry:
> 
> - primary antibody incubation overnight at 4 degrees
> ?
> 
> - primary antibody incubation 30-60 min at room
> temperature ?
> 
> With compliments
> 
> * * * * * * * * * * * * * * * * * * * * * * * * *
> Fernando Capela e Silva
> Laborat#243#rio de Biologia da Conserva#231##227#o
> Departamento de Biologia
> Universidade de ...vora
> Apartado 94
> 7002-554 ...vora
> PORTUGAL
> 
> Phone: +351-266 760 800
> Fax: +351-266 711 231
> Email: fcs@uevora.pt
> 
> http://evunix.uevora.pt/~fcs/FernandoCapelaSilva.htm
> 
> 
> 


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