If these smears are just fixed in acetone you could probably eliminate
the retrieval portion of your protocol. I would also block for endog.
> Hello fellow Histonetters:
> We have started experimenting with Zymed's TRAcP antibody with
> poor results on peripherial smears. Does anyone have any guidelines
> to follow to bring this antibody up and running? Washing off is a
> problem as well.
> 1. Fix in Acetone for 1 minute.
> 2. Steam retrieval in EDTA for 30 minutes.
> 3. Cool for 30 minutes.
> 4. Primary antibody for 60 minutes.
> 5. Link for 30 minutes (LSAB2)
> 6. Strep for 30 minutes.
> 7. DAB for 5 minutes
> 8. Counterstain with Hema for 5 - 8 seconds.
> The procedure is run on DAKO Autostainer.
> Thanks everyone for your help.
> Jim Bradford
> Orlando, FL
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