Re: how to dehydrate slides stained with AEC

From:"#177##163#OE#192#"

Dear Kiernan,
Thank you for your advise. 
I had modified my experimental strategy. I used mouse spleen to test the system since I was intended to indentify CD4 cell. I did it as following on acetone fixed frozen section:
(1) negative control was done with primary antibody being omitted
(2) positive control was included too.
(3) primary antibody, 30 min
(4) 2nd antibody, 30 min
(5) Enzyme, 30 min
(6) AEC substrate, 6 min
(7) hematoxylin conterstain, 5 min
There was red stain on negative control slide but not all the silde while the positive was all stained red very where. I noticed the 2nd antibody was antimouse. But my primary antibody was take from rat. Must I use 2nd anti-rat antiboy? 

 Baowei Peng
 Shanghai JiaoTong University,
 Shanghai,200030
 PRC

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