[Histonet] PECAM-1 (sc-1506R) staining
vhlwong <@t> yahoo.com
Wed Jul 28 23:05:45 CDT 2010
Thank you for all the professional suggestion of the previous IHC staining and finally I got some positive results.
Then comes problem on the PECAM-1 IHC. I am using the Santa Cruz sc-1506R (it is a anti-RAT antibody from RABBIT). I stain decalcified rat spine paraffin sections, with following steps:
antigen retrieval (~95C MW, DAKO AR solution pH6 for 20 minutes);
DAKO's Dual Enzyme block for 10 minutes;
3% normal goat serum for 20 minutes;
1:50 primary antibody at 4C for overnight;
DAKO's Envision anti-rabbit kit for 30 minutes;
DAKO's DAB solution to develop
I used rat liver and kidney as positive controls. I got background staining on hepatocytes but none staining on vessels. On kidney section, nucleus and lining within tubules and basement membrane of glomerulus were also stained but not that bad as liver. I cannot say it was totally non-specific as some nucleus were not stained.
I think PECAM-1 IHC needs more specific conditons. Could any histoneters have any suggestion?
Many thanks in advance.
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