Roger and all,
The main component of the Pierce Neutral Elution buffer is magnesium
chloride.
According to my former colleagues in Protein Chemistry at Amgen you can make
a near-equivalent with 3.6 M magnesium chloride, 0.2 M sodium acetate, pH
6.55. I think the real Pierce stuff contains a bit of detergent also.
John Philo
Alliance Protein Laboratories
www.ap-lab.com
-----Original Message-----
From: Association of Biomolecular Resource Facilities
[mailto:abrf-request@aecom.yu.edu]On Behalf Of gsarath@unlnotes.unl.edu
Sent: Wednesday, June 14, 2000 7:13 AM
To: Recipients of ABRF List
Subject: Re: "gentle elution buffer"
Hi Roger: I have been using the gentle elution buffer for a while. It
works like a charm. I am uncertain what salt causes precipitation to
occur. Last month, we had a missed communication and the student dialysed
the eluted fraction from an ligand-affinity column into PBS. There was a
lot of precipitate, and I thought that all was over and the skies would
fall, but NO! We spun out the precipitate and the eluted antibodies were
still present, although I m not sure as to how much we lost. When we use
TBS for dialysis, we have faced no problems and purified Ab titer is very
good.
I have essentially switched over to the Pierce Gentle Elution Buffer for
all our Ab-affinity purification protocols. My suscpision would be that
they have a large amount of titrating Cation that precipitates out as the
phosphate salt. Take your pick, Gautam
Gautam Sarath
N-226, The Beadle Center, 19th and Vine Street
University of Nebraska, Lincoln
Lincoln, NE 68588-0664
Tel: 1-402-472-2928; Fax: 1-402-472-7842
http://www.biotech.unl.edu/Proteins/index.html
http://www.pigment.unl.edu/dept/sarath/sarath.html
This archive was generated by hypermail 2b29 : Fri Jun 30 2000 - 07:51:55 EDT