Dear All:
Just wonder anyone have worked on PSA_NCAM IHC of frozen sections?
How about 1:1000 mouse anti-PSA NCAM from millipore to stain 30 um brain sections?
And, do you need to add triton into the PBS, to "pore" the membrane?
I think PSA-NCAM is distributed on the cell membrane, thus PBS may work?
Protocol:
1. Rehydration. PBS
2. Mouse-anti PSA-NCAM 1:1000 overnight + Blocking simultaneously.
3. 3* PBS
4. Goat-anti mouse conjugated with 488 fluroscein
5. 3 * PBS
6. DAPI
7. PBS wash
8. mount
Any other comments on this?
especially the time & concentration for first antibody?
thx,
2008-07-31
tf
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