Re: [Histonet] troubles with CD4

From:Lori Richey

We were having alot of problems with CD4.
We use  NOVACASTRA CD4 . The data sheet suggested using 0.5% hydrogen 
peroxide in Methanol for 10 minutes to quench.  This  along with 15 
minutes microwave in EDTA Ph 8 has been working well in our system.

Thomas Pier wrote:

>Hello all,
>I was wondering if anyone could help me with some problems that I'm having with staining for CD4.  My control tissue for the work up is formalin fixted, paraffin embeded tonsil.  I'm using Labvision's CD4 Ab-2 clone 1F6.  I have diluted it anywhere from 1:10-1:50.  I'm using 1% goat serum in TBS as my diluent.  I've tried EDTA HIER as recomended on the data sheet.  I've also tried protease XXV w/ or w/o the EDTA.  I shouldn't be running into the peroxidase problem since I'm using an alkaline phosphatase detection system (Biocare's Vulcan Fast Red).  I have also tried using or not using a 10% goat serum protein block.  I have gotten some staining, but nowhere near as much as I should in a tonsil.  Any help that you could give me would be greatly appreciated.
>Tom Pier
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