RE: IHC in BM cytospins!

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From:"Nocito, Joseph" <>
To:"''" <>,

we normally fix our BM cytospins in cold methanol for 20 minutes.  As
always, there are exceptions.  We found out the myeloperoxidase stains
better in Acetone.  In either case we block for endogenous peroxide for 15
minutes, protein block for 5 minutes and have a 30 minute incubation of
30-primary antibody, 30-link, 30-label and 5 minute DAB.  We use Dako's
LSAB2 kits with good results.  Hope this helps

Joe Nocito, B.S., HT(ASCP)QIHC
Histology Supervisor
Christus Santa Rosa Hospitals
San Antonio, Texas 

> -----Original Message-----
> From:
> []
> Sent:	Monday, February 07, 2000 8:21 AM
> To:
> Subject:	IHC in BM cytospins!
>      Greetings to All!  I have a question concerning IHC in bone marrow
> cytospins.  Could anyone please recommend an appropriate fixative, time
> and
> conditions?  Also, are there any "tricks of the trade" or specials
> conditions
> that don't apply to IHC in frozen tissue???? I am trying (STILL) to
> immunostain
> for CD45 in rat bone marrow and spleen cytospins with a varying degree of
> success!  Any suggestions would be very much appreciated!  Thank you as
> always!
> Jennifer Hoover
> Novartis Pharmaceuticals

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