|From:||Kim Kusser <firstname.lastname@example.org>|
I stain all the time with PNA on mouse spleen. I suppose I can just tell you what I do:
1. cut frozen sections 5-7um-----> air dry (1hr to O/N)
2. Fix in 4c acetone for 10 minutes -----> air dry ---> PBS
3. Block endogenous peroxidase with 0.1M NaN3 for 10 minutes ----> PBS 3X (3x 5 minutes)
4. Serum block for 30 minutes (I use 5%BSA in dPBS)
5. Avidin block 15 minutes ----> pBS1X ----> Biotin block 15 minutes ----> PBS 2X
6. PNA-b 1:800 for 30 minutes ------> PBS 3X
7. ABC (from vector) for 30 minutes ----> PBS 3X
8. Develope with DAB
PNA is very sticky, so you'll want to make sure you titrate it as high as you can. But I routinely get very nice germinal center staining.
Hope this helps.
Date: 1 May 2001 16:49:33 -0500
Subject: PNA staining
We are having difficulty get nice PNA (for Germinal center B cells)
staining on mouse spleen and LN. Can any one share some experience about
this staining with us? Thanks!
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