I agree with you about the current limitations of the herceptest protocol. And
as you indicate, the effectiveness of target retrieval is based on the
temperature achieved. I know that CAP is circulating a kit to different
insititutions for comparative validation and this might be a problem they would
like to address.
Louise, I know this is of not much help to you but ...
Abizar
www.innogenex.com
1-877-IGX-INFO

Louise Taylor wrote:

> Dear Abizar
>
> This may be the case, BUT, I work at 6000ft above sea level, where boiling
> temp is 93.5 deg C so unless i pressure cook - I cannot acchieve the boiling
> temp of 95-100. So whats the option?
>
> Regards
> louise
>
> -----Original Message-----
> From: Abizar Lakdawalla [mailto:abizarl@innogenex.com]
> Sent: 05 September 2000 09:12
> To: Sebree Linda A.
> Cc: 'Louise Taylor'; 'Histonet'
> Subject: Re: Hercep help
>
> Just a comment: Changes in protocol for FDA tests are usually not allowed.
> You
> have to follow the intructions to the dot.
> abizar
>
> "Sebree Linda A." wrote:
>
> > Hi Louise,
> >
> > When we use the waterbath for HIER, we first bring the buffer and slides
> (in
> > a plastic coplin jar) up to a near boil in a microwave oven and then
> > transfer it to the waterbath and start our timing.  This works very well
> for
> > us and is our most gentle form of HIER that we use.
> >
> > Linda A. Sebree, HT
> > University of Wisconsin Hospital & Clinics
> > Immunohistochemistry/In Situ Hybridization Laboratory
> > D4/218-2472
> > 600 Highland Avenue
> > Madison, WI  53792-2472
> >
> > (608)265-6596
> > FAX: (608)263-1568
> >
> > > -----Original Message-----
> > > From: Louise Taylor [SMTP:louiset@mail.saimr.wits.ac.za]
> > > Sent: Monday, September 04, 2000 10:08 AM
> > > To:   histonet
> > > Subject:      Hercep help
> > >
> > > HI histonetters
> > >
> > > I need some help urgently!  I am embarking on  a  hercep trial and have
> > > run
> > > into an unexpected problem. What do you use to put your slides in to
> > > "boil"
> > > them for antigen retrieval? I tried using a glass coplin jar in my
> > > waterbath, but it shattered, and when I used a plastic one, I noticed
> that
> > > the internal solution temperature did not go above 82 degrees. (I have
> > > never
> > > been required to use a waterbath for Ag retrieval, having quite happily
> > > used
> > > a slide rack and a beaker in the microwave - but this sytem won't really
> > > work, as it uses quite a lot of buffer).
> > >
> > > Any suggestions will be welcome, I have about 100 slides to do in the
> next
> > > few days before I go on honeymoon!!
> > >
> > > Many thanks
> > >
> > > Louise Taylor
> > >
> > >
> > > Research Laboratory
> > > Department of Anatomical Pathology
> > > South African Institute for Medical Research
> > > Johannesburg
> > > South Africa
> > >
> > >
> > >