[Histonet] autoclave vs. filter of RBC lysis buffer

Oble, Darryl A.,M.D. DOBLE <@t> PARTNERS.ORG
Sun Jun 4 14:23:49 CDT 2006

I was wondering whether anyone knew why RBC lysis buffer is always filtered and
not autoclaved. Below are the ingredients. I saw some material on the internet
about corrosion of the autoclave, but this was for a different NH4Cl containing
solution. Thanks for any help that you can offer.

4.14g NH4Cl
0.5g KHCO3
0.018g EDTA
dissolve in water
adjust pH to 7.35 with 0.1NaOH
Total volume 500ml

Frustrated Scientist

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Today's Topics:

   1. (no subject) (Heike Grabsch)
   2. Re: section adhesion on slides (tiang yeepeng)
   3. re: Bulk DAB (Carl Hobbs)


Message: 1
Date: Sun, 4 Jun 2006 11:48:54 +0100
From: "Heike Grabsch" <H.I.Grabsch <@t> leeds.ac.uk>
Subject: [Histonet] (no subject)
To: <quinntl <@t> umkc.edu>
Cc: histonet <@t> lists.utsouthwestern.edu
	<C6AC304539F43C4196253FD94B7960F316A81A <@t> HERMES2.ds.leeds.ac.uk>
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Dear Tim, 
ScyTek in the US do sell bulk reagents for immunohisotchemsitry which are more
or less ready to use. On the other hand you may consider going back to the 'good
old days' where we used to make up the DAB from either from actual DAB powder or
Dr Heike Grabsch, MRCPath MD
Gastrointestinal Research Group
Pathology and Tumour Biology
Leeds Institute of Molecular Medicine
St James's University Hospital 
JIF Building, Level 4
Beckett Street
Date: Fri, 2 Jun 2006 12:22:45 -0500
From: "Quinn, Tim L." <quinntl <@t> umkc.edu>
Subject: [Histonet] Bulk DAB
To: <histonet <@t> lists.utsouthwestern.edu>
        <DF0E3D9A75932A458B25C1889377445A01C9C472 <@t> KC-MSX4.kc.umkc.edu>
Content-Type: text/plain;       charset="us-ascii"

Dear Listies,

I would like to start using 250 mL tubs for DAB staining to get staining
time standardized. Is there a source for large purchase of reagents at
an affordable price?


Timothy L. Quinn
Senior Lab Technician
Missouri University at Kansas City
Medical School- Med Lab
2411 Holmes
Kansas City, MO  64108
quinntl <@t> umkc.edu



Message: 2
Date: Sun, 4 Jun 2006 15:47:07 +0000
From: "tiang yeepeng" <yeepengtiang <@t> hotmail.com>
Subject: Re: [Histonet] section adhesion on slides
To: <histonet <@t> lists.utsouthwestern.edu>, <mcauliff <@t> umdnj.edu>
Message-ID: <BAY108-W89CBD65E8611269556E75DF970 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"

I coat my slides with poly-L-lysine but it seemed that I lost many cells during
the washing steps in IHC. I use a centrifuge my cells onto the slides using the
cytospin. Any idea or comment, Geoff?

With regards,
Department of Medical Microbiology,
Faculty of Medicine,
University of Malaya.
It's the future, it's here, and it's free: Windows Live Mail beta


Message: 3
Date: Sun, 4 Jun 2006 17:37:16 +0100
From: "Carl Hobbs" <carl.hobbs <@t> kcl.ac.uk>
Subject: [Histonet] re: Bulk DAB
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <DIEOLOLFLGEINEHLAHFEKECICCAA.carl.hobbs <@t> kcl.ac.uk>
Content-Type: text/plain;	charset="iso-8859-1"

My recipe here: http://www.immunoportal.com/index.php Great site: not mine
but I contribute. I must admit that I no longer heat before adding water to
DAB. It dissolves well in RT water.
Hope that's helpful


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