[Histonet] RE: Antigen retrieval

[Mark Tarango]

Keep in mind that the PT module can't get to above 100 C without boiling
over.  For some antigens you need to get it hotter than 100 C.

On Sun, Apr 13, 2008 at 11:51 PM, C.M. van der Loos <
c.m.vanderloos <@t> amc.uva.nl> wrote:

> Kim,How about using a Thermo PTModule for this? You can even program this
> thing for overnight applications! In the past we used a stove for overnight
> at 70C. First, a tank with HIER buffer was warmed up in the microwave and
> then placed in a stove (70C was the maximum setting for it) with a glass
> plate covering the tank. The effect of overnight antigen retrieval at 70C
> for the few antigens we tested was a bit different, but mostly comparable
> with the effect of HIER for 20 min at 98C. Some of the antigens responded
> very well and showed far more intense staining than after short HIER (20
> min, 98C), and others responded very poor. The big advantage of overnight
> HIER at 70C was that sections from fatty tissue better stayed on the glass
> slides.Hope this info helps a bit!Cheers, ChrisChris van der Loos, PhD
> Dept. of Pathology
> Academic Medical Center M2-230
> Meibergdreef 9
> NL-1105 AZ Amsterdam
> The Netherlands Date: Sat, 12 Apr 2008 17:18:10 +1000
> From: Kim O'Sullivan <Kim.Osullivan <@t> med.monash.edu.au>
> Subject: [Histonet] Antigen retrieval
> To: histonet <@t> lists.utsouthwestern.edu
>
> Hi,
>
> Is anyone familiar with doing antigen retrieval overnight at 80 degrees
> celsius in 0.1M Tris/HCL pH 9.0? My question is: What is the best way to do
> this in terms of safety etc. A waterbath? Hot oven? Any idea's appreciated.
>
> Kim O'Sullivan
> Monash University
> Melbourne
> Australia
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