[Histonet] Pronase antigen retrieval

[Hoekert, W.E.J.]

Hello Eva,
 
We are not using pronase anymore, but when we did, we were doing it at room temperature. (15 mg in 250 ml PBS, sigma P-5147). It worked fine. 
 
Our Trypsin antigen retrieval, we were doing it at 37C as follows: the evening before the retrieval, we put some PBS in the 37C stove, the next morning we dissolved the trypsin in the warm PBS and we put the slides in it (again in the stove, for 15 minutes). 
 
Good luck,
 
Willem

________________________________

Van: histonet-bounces <@t> lists.utsouthwestern.edu namens Eva Permaul
Verzonden: di 11-9-2012 19:36
Aan: histonet <@t> lists.utsouthwestern.edu
Onderwerp: [Histonet] Pronase antigen retrieval



Hello out there in Histoland,
I have to do some bulk runs with Pronase antigen retrieval. Up until now
the number of slides have been so few that I have been able to do the
antigen retrieval by dropping pronase on each slide and incubating them in
a humidified chamber at 37C for 20min. I now have so many slides that my
manager has suggested that I make up large amounts of the Pronase (250ml)
and submerge the slides. She suggested placing the container in the oven
earlier to allow the Pronase to get to 37C before I add the slides. How
long is the Pronase ok to be keep at 37C before I use it? How long in
advance can I place it in the 37C before it looses its ability to be used
as an antigen retrieval?
Thanks for all your help,
Eva Permaul
HTSR
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