RE: Dako autostainer

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
To:"Greer, Patricia" <pwg1@cdc.gov>, histonet@pathology.swmed.edu
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By trash in your swine serum, do you mean protein aggragates floating
around??  If so, you can use a microfilter (they come as Millipore
Steriflip, 50 ml disposable vacuum filtration system, large centrifuge tube
in 0.22 um pore size and probably 0.45 um).  This cleans the floating
garbage, filter sterilizes your serum and leaves it clean, clear and ready
to use.  There are syringe filters also.  We use these for both heat
inactivated or thawed serums/reconstituted lyophilized serums with
excellent results.  You can also microfilter the 20% serum after you make
that up.      

At 01:09 PM 7/24/00 -0400, you wrote:
>Denise,
>We use Tris buffered saline with Tween and magnesium chloride for our buffer
>washes.  It is not as convenient as the Dako buffer and it has to be
>refrigerated to prevent bacterial contamination.  This is the same buffer we
>used when we were doing our immunos by hand and preferred not to change.
>For antibody diluent we use 20% normal sheep or swine serum in the wash 
>buffer(this is the same solution we use for protein blocking).  Normal swine
>serum is preferred since that is the species of the link, but we were having
>a lot of trouble with "trash" in the swine serum.
>
>Pat Greer
>CDC
>Infectious Disease Pathology 
>
> -----Original Message-----
>From: denise M m Long-Woodward [mailto:denisew2@juno.com]
>Sent: Monday, July 24, 2000 6:03 AM
>To: histonet@pathology.swmed.edu
>Subject: Dako autostainer
>
>
>I've been asked to relay a question to all those who work with Dako's
>Autoimmunostainer. What are you using for Buffer rinses: Dako's
>commercial  TBS or something else?  Those who use something else, could
>you please tell me what it is?   They also want to know what you are
>using to dilute your antibodies. Are you using Dako's commerical stuff?
>Again please specify the diluent being used. 
>
>Many thanks for your help!
>
>Denise Long Woodward
>________________________________________________________________
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>
>
Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303



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