Re: Cox-1 and Cox-2

From:r-meyer2@northwestern.edu



Dana, 

I have worked with the Cox 2 monoclonal mouse antibody on FFPE human tissue 
from Cayman Chemical, Ann Arbor, MI (Cat. # 160112; 800-364-9897).  I used 
colon adenocarcinoma as a control with a dilution of 1:100.  For pretreatment, 
I used citrate buffer pH 6.0 (DAKO 10X Target Retrieval Solution, S1699) for 20 
minutes in a steamer when the buffer had reached above 95 degrees celsius, and  
a 20 minute cool down at room temperature.  I used DAKO EnVision + detection 
system -- peroxidase block (DAKO S2001) for 10 minutes, protein block (X0909) 
for five minutes, primary ab for 30 minutes, anti-mouse secondary ab/HRP 
polymer for 15 minutes, and DAB/H2O2 for 7 minutes.  The staining is excellent 
and everyone here was really happy with the results. 

Bob Meyer, HTL 
Pathology Core Facility 
Northwestern University 


Dana Settembre  writes on Thu, 11 Apr 2002 15:37:28 -0400: 
> 
> 
> Hello, 
> I'd like to run Cox-1 and Cox-2 on FFPE human tissue.  Can't seem to get it 
to work. 
> Which vendor do you use?  What about control? Pretreatment?  Dilution?   
Help! 
> 
> Thank you, 
> Dana Settembre 
> University Hospital-UMDNJ 
> Newark, NJ 
> 
> 
> 








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