Re: CD3 has too much background, ca you help?

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From:Dana Settembre <settembr@UMDNJ.EDU> (by way of Marvin Hanna)
To:histonet@histosearch.com
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On Thu, 27 Jan 2000, Sandy Julsing wrote:

> Histo-buddies
>      Our CD3 has too much background stain including in the gastric
>glands. Our protocol is...
>
> -DAKO pre-dilute CD3
> -antigen retrieval with citrate buffer pH 5.8-6.2 for 20 min with a 20
>min cool down
> -LSAB2 detection kit 10min/10min/5min
> -Stained on the DAKO autostainer
>
>  Any help would be greatly appreciated, thanks,  Sandosis
>
>

Hello Sandy,
I use Dako's polyclonal CD3 and I pretreat for 5 minutes room
temperature in the autostainer.  I use Dako's code no. A0452 at a dilution
of 1:100 with their prot. K code S3020 on a tonsil positive control. Very
nice results.  (I also use their LSAB2 kit).  Good Luck!

Dana Settembre
Immunohistochemistry Lab
Pathology Department
University Hospital
Newark,  New Jersey
USA




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