Re: Hercep help

From:Amos Brooks <> (by way of histonet)

    Congratulations and best wishes on your honey moon!
    I too grumbled about the water bath. As I understand it, this is what the
FDA approved and since that is what they agreed to, they are very tight about
it. Other methods will work but they are not backed up by the FDA, and the labs
cannot claim they are. The FDA is just trying to eliminate lab to lab
Which, believe me, I see happens often.
    We have always used a water bath incubator. the Fisher and CMS catalogs
good ones that describe the volume of water they use, and the maximum and
minimum temperatures they maintain. Most also have a digital display of the
water temperature. The plastic lids often have a hole in them to hold a
thermometer (those that don't a hole can be easily drilled into the Plexiglas
top to accommodate one). I would highly recommend one of these.
    As for the coplin jar, avoid the glass ones, but you already found that out
(a day late and about $20 short right). I have had good luck with plastic
jars and plastic staining dishes with a slide rack. I have noticed that the
escapes if you have the lid ajar to check the temperature. That is why I like
the idea of a lid with a thermometer sized hole in it, there's less of a heat
sink. if you need more help feel free to contact me.

best of luck
Amos Brooks
    ps: By the way Andi Kappeler's bunsen burner idea scares the heck out of me
not so much because of the open flame (not that that's not an issue) but
I don't think the heat would be distributed evenly, and there could be many
temperature fluctuations in the process of regulating the temperature. (sorry
Andi, I just don't like it, but that is just me)

Louise Taylor wrote:

> 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

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