AW: [Histonet] Trypsin versus other AR-s

[Gudrun Lang]

If PIER is needed, we use Proteinase K, but we avoid it, if possible.
Citrate Buffer is historically the next buffer, that got generally used. Few
years later with investigations about Ca-Ions involved in fixation, the
EDTA-buffers with high pH appeared.
It was shown, that most of the diagnostic antibodies work well with high pH
retrieval. So my first choice is the EDTA-buffer when introducing a new
antibody. Although the most datasheets require Citrate buffer.
Gudrun  

-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Margaryan,
Naira
Gesendet: Freitag, 27. März 2009 20:38
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] Trypsin versus other AR-s

Hi Dears,

 

It is Friday, but not a weekend yet:-):-(

 

I just got a request from my PI to figure out:

1.      How often now day's people use Trypsin (EDTA, Proteinase K or E)
as an Antigen Retrieval for FFPE. 

2.      Why or is the Citrate Buffer pH6 more usable??? 

3.      Is Trypsin very old technique? 

 

Any educational feedback is appreciated,

Naira

 

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