RE: [Histonet] EDTA pH 8.0 recipe

From:"Bonnie Whitaker"

Did I miss something?  I assumed that she wanted it for HIER.... maybe my
prejudice is showing, but that's what I use EDTA at pH 8 for.

Bonnie Whitaker
-----Original Message-----
[] On Behalf Of Gayle Callis
Sent: Wednesday, March 24, 2004 10:54 AM
To: Barnhart, Tammy;
Subject: Re: [Histonet] EDTA pH 8.0 recipe

Use EDTA Tetrasodium salt, dissolve in 800 mls distilled water and adjust
pH DOWN to pH 8 using pH meter and glacial acetic acid, just titrate it
with a pipette, watching the meter.  This is an old method from Webb Gee
from U of Utah.  The pH of tetrasodium EDTA is around 9 to begin with.  If
you are planning to decalcify bone with pH 8, remember this is rather
alkaline and protein linkages sensitive to alkaline conditions can be
damaged.  Maybe work at pH 7.4 - 7.6, and stay out of higher pH range.  

You can dissolve as much as 14 gm/100 mls buffer or distilled water.  IF
the bone is totally fixed to begin with, distilled water can be used to
make up the EDTA solution. Tetrasodium EDTA goes into solution much faster
than EDTA, or EDTA Disodium salt (they are soluble to around 10% aka 10
g/100 mls).  

Good luck

At 07:57 AM 3/24/2004 -0600, you wrote:
>Does anyone have a recipe to make EDTA buffer at a pH of 8.0.  I would like
>to make our own in an effort to save some dollars.  I have checked the
>archives and only have come up with one rather difficult procedure. Thanks
>in advance....
>Tammy Barnhart, BS, HTL(ASCP)
>Anatomic Pathology Supervisor
>St. Alexius Medical Center
>Bismarck, ND
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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