Re: Method for double immuno staining

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From:amos brooks <>
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    Contact a reputable vendor (DAKO, Ventana, etc) and ask them for a double
staining kit. This should come with relatively easy to follow directions.
    You will only need to epitope retrieve once. The crosslinked proteins once
broken remain so thruought the staining procedure. One peroxidase block should do as
    The newer procedures are growing more and more easy to follow. You may be able
to get them to send you the packing slips to review so you can compare the
Amos Brooks wrote:

> A doctor has asked our lab to do a double immuno stain using both p21 and ki 67.
> My question is---where can I look for a relatively easy to understand method to
> accomplish this?  I suspect that I will need to use 2 different chromagens(dab
> and aec) but do I have to retreive the antigens twice, block peroxidase twice,
> etc?
> My assumption is that I should proceed with one antibody stain form start to
> finish and then for the second antibody I should start the procedure from the
> point of primary antibody application to the second chromagen.
> Any help anybody can give regarding either the method or where to look up the
> method would be greatly appreciated.
> Sincerely,
> Christine Klein
> MD Anderson Cancer Center
> Houston, Tx.

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