[Histonet] Antigen retrieval

[Kim Merriam]

Hi Kim,
   
  I have done overnight retreival on a few occasions (although it was at 60-65 and not 80), wtih very good success.  I have done it in an oven, with the lids from the tissue tek plastic dishes taped on or covered in foil.  It probably does not matter if it is an oven or a waterbath, as long as you are confident that the temperature can be maintained properly.
   
  Another note, I have also done this with high-pH retreival (10-10.5 or so) solution, which is fine as long as the tissues are not too fatty, otherwise they can not survive such long times at that pH. 
   
  Good luck,
  Kim Merriam
   
  
Kim O'Sullivan <Kim.Osullivan <@t> med.monash.edu.au> wrote:
  Hi Gayle, Renee and others,

Thanks for the reply's. I am staining for KIM-1/TIM-1 and all the literature states that it has to be subjected to antigen retrieval at 80 degree's celsius overnight,- I was just worried that I would come back to the lab the next day to find either the lab had burned down or my tissue ruined!

Kim 

Gayle Callis wrote:> 
> Rene and Kim,
> 
> If you cover the waterbath (they come with covers) and also the slide
> container (coplin jars, whatever), it should eliminate evaporation.
> 
> Waterbath methods are also found in the literature. Jules Elias has
> published a method but I do not recall the temperature used overnight.
> 
> I have several waterbath heated methods that use higher temperatures for
> shorter times if Kim is interested. I will be happy to provide those via
> private message with attachment.
> 
> Gayle M. Callis
> HTL/HT/MT(ASCP)
> Bozeman MT 59715
> 
> 
> ----- Original Message -----
> From: "Rene J Buesa" 
> To: "Kim O'Sullivan" ;
> 
> Sent: Saturday, April 12, 2008 9:03 AM
> Subject: Re: [Histonet] Antigen retrieval
> 
> 
>> Never heard of such a procedure. On the other hand, unless you are
>> dealing
>> with a HUGE amount of buffer, it will evaporate during such a
>> prolonged
>> period of time.
>> Check your reference, I think something is not right.
>> René J.
>>
>> Kim O'Sullivan wrote:
>>
>>
>>
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Kim Merriam, MA, HT(ASCP)
Cambridge, MA