Re: ki-67 protocol

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From:Katri Tuomala <katri@istar.ca> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Daisey Joseph wrote:
>
> Hi Histonetters:
> We have tried everything from different pretreatment protocols to
>different concentrations of the antibody and have concluded that antigen
>Ki67 antigen is masked by the Prefer fixative.  I have been in contact
>with other labs using Prefer fixative and the experience is mutual.   If
>someone out there has found  a way to make it work, I'd like to know also.
>
> Daisy Joseph
> Impath  Inc.
>
> -----Original Message-----
> From:   Kathy-Jean Carney [SMTP:carnek@dianon.com]
> Sent:   Tuesday, November 10, 1998 7:16 AM
> To:     'Histonetters'
> Subject:        ki-67 protocol
>
> Does anyone have a protocol for Ki-67 (Zymed or Immunotech antibodies)
>when tissue is fixed in Prefer?  Our Immuno's are done by automation.  We
>do not antigen retrieve with prefer, but is there something else we can
>do?  We are getting all negative staining and I know some should be
>positive. Thanks
> Kathy
Hi there!
It seems to me that this Prefer fixative simply does not preserve this
particular antigen. I don't know what is in Prefer (trade secret), but
many so called formalin substitutes are alcohol based and some antigenic
epitopes are not preserved by alcohol.
Just a thought! Katri.




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