Patsy,
I'm sort of coming in on the middle of this so I hope
I don't make a major gaff here.

When doing the long term HIER take your protocol up to
the primary antibody.  Apply the primary and cover the
slides with strips of parafilm.  Then incubate the
slides overnight at 4ºC in a sealed humid chamber. 
Next day you can complete the balance of your
protocol.  I've done with with a variety of ab's on a
variety of species. 

Hope this helps.

Vikki Baker
Mt. Sinai School of Medicine 
--- Jim Staruk  wrote:

> If I remember correctly, mine was an anti-collagen I
> or II that did not work
> after sitting in buffer overnight.  I also know that
> one of my part-timers
> tried this at the hospital he works at and they also
> had disastrous results
> on day #2.
> 
> ______________________
>     Jim Staruk
> Mass Histology Service
> www.masshistology.com
> 
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu]
> On Behalf Of Kim Merriam
> Sent: Friday, February 18, 2005 7:42 AM
> To: Jim Staruk; 'Patsy Ruegg';
> ihcrg@neo.agsci.colostate.edu
> Cc: histonet@pathology.swmed.edu
> Subject: RE: [Histonet] antigen retrieval question
> 
> Patsy,
>  
> I have done this many times before, and have never
> had any issues with
> re-masking.  Why would the sites be re-masked if
> they are not subjected to
> formalin again?  If I am staining a big batch of
> slides (anything over 100
> slides at a time), I do the depar and retrieval on
> the first day and all of
> the antibody stuff on the second day.  Maybe it just
> depends on the
> protein/antibody?
>  
> This has peaked my interest.  I am curious to see if
> anyone else has
> encountered this problem.
>  
> Kim Merriam
> Novartis
> Cambridge, MA
> 
> 
> 
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> Histonet@lists.utsouthwestern.edu
>
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 



		
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