Re: citrate buffer recipe

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From:Jochen Schuck <jschuck@otogene.de>
To:histonet@pathology.swmed.edu
Reply-To:
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Hi Linda,
we use the following recipe for a 10mM citrate buffer at pH 6,0:

stock solution A (0,1M citric acid): 21,01g C6H8O7 x 2H2O in 1000ml aqua
dest.
stock solution B (0,1M sodium citrate): 29,41g C6H5O7 x 2H2O in 1000ml aqua
dest.

For the buffer take 9ml solution A and 41ml solution B and add 450ml aqua
dest. We use this buffer for microwave antigen retrieval: we put the slides
into a cuvette (after deparaffination) with this buffer and heat at full
power in the microwave until the buffer boils violently. Then we take 5 min
at full power. Make shure you have something to cover the cuvette, otherwise
your concentrations will change drastically. After cooking we let the
sections cool down to approx. 40C. Then comes the staining.
Good luck and happy staining!

Jochen Schuck

P.S.: In order not to loose your tissue during the cooking process you need
really good adherence of the tissue to the slides....

> Von: "Sebree Linda A." <la.sebree@hosp.wisc.edu>
> Antworten an: histonet@pathology.swmed.edu
> Datum: Mon, 27 Mar 2000 14:42:08 -0600
> An: 'Histonet' <histonet@pathology.swmed.edu>, 'IPOX'
> <ipox-l@pathology.stanford.edu>
> Betreff: citrate buffer recipe
> 
> Our lab would like to make its own citrate buffer for HIER purposes; if
> anyone out there has a "tried and true" recipe for such, we would appreciate
> hearing from you.
> 
> Thanks in advance,
> 
> Linda Sebree
> 
> 
> 
> 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
> 
> 




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