Hi Guoji,

Thank you for your input!  Yes, I'd love to have the reprint.  Could you
send it to:

Mari Shinohara
Department of Immunology and Immunogenetics
Joslin Diabetes Center
Boston, MA 02215

Thanks a million.

Mari

-----Original Message-----
From: Wanda Wang [mailto:[log in to unmask]]
Sent: Thursday, August 24, 2000 12:55 AM
To: [log in to unmask]
Subject: Re: triple step staining


Hi! Mari:

    We have a paper on METHODS 18, 459-464 (1999) "Tyramide Signal
Amplification Method in Multiplelabel immunofluorescence Confocal
Microscopy". Please let me know if you need a reprint of this paper (send
you mail address to me).


    Good luck,

    Guoji Wang



----------
>From: "Shinohara, Mari" <[log in to unmask]>
>To: [log in to unmask]
>Newsgroups: bit.listserv.confocal
>Subject: triple step staining
>Date: Thu, Aug 24, 2000, 11:11 PM
>

> Hello,
>
> Since I'd like to detect a protein existing at very low levels, I'm
thinking
> to try triple step staining: using a primary Ab, a secondary Ab, then a
> fluorochrome conjugated tertiary Ab.  No one around me have done it and I
do
> not have any idea.  Any suggestions will be greatly appreciated regarding
to
> especially the following questions...
>
> (1) The protein is intracellular (ER and Golgi).  I wonder whether the
> bulkiness of the whole molecules affects staining efficiency...?
>
> (2) Do you know any reference which helps for the triple staining?
> Anything, papers/protocols/books, will be welcome.
>
> (3) Or are there any good alternatives, rather than doing the triple
> staining, to detect low level proteins?
>
> Thank you very much in advance.
>
> Mari
>