Thanks Derek, I will definitely pass your comments on to the interested
parties.
Helen Kim, Ph.D.
Res. Associate Professor
Department of Pharmacology & Toxicology
Director, 2-D Proteomics Laboratory
University of Alabama at Birmingham
1670 University Blvd, Volker Hall, G78P
Birmingham, AL 35294-0019
(205) 934-3880 (office, with voicemail)/5-3244 (lab, no voicemail)
FAX: (205) 934-6944
helenkim@uab.edu
> ----------
> From: Derek Van Dyk
> Sent: Sunday, February 18, 2001 4:35 PM
> To: Recipients of ABRF List
> Subject: Re: Blue-Sepharose
>
> Helen
>
> we have used Cibacron Blue ( Sigma) to deplete albumin prior to 2 DE with
> a
> certain amount of success: the process certainly does not remove all
> albumin but it reduces it to the extent that there is an increase in the
> resolution of other less abundant proteins.
>
> Suggested buffers are 50mM Phosphate, neutral pH or We use 50mM Tris, 50mM
>
> NaCl, pH 7.2. There are some reports that suggest that tris shows
> decreased
> albumin binding relative to phosphate, so you might want to try 50mM
> phosphate.
>
> Depletion can be done either by batch binding in a microfuge tube or by
> packing a small column in a pipette tip. Bed:sample ratio needs to be as
> large as possible: suggest 3:1 v:v
>
> Something to be aware of: there is a certain amount of non specific
> binding
> to other plasma proteins.
>
> Good luck
>
> Derek
>
>
>
>
> At 16:42 16/02/01 -0600, Helen Kim, Ph.D. wrote:
> >Hi All,
> > Has anyone had experience removing albumin with Blue-sepharose?
> I
> >know in principle it can be done, and I've read lots of papers where
> people
> >blithely say they did with the blue sepharose. I recommended the
> >procedure to a grad student dealing with blob of albumin in his 2-D
> gels
> >of conditioned media. He bought some, tried it (I don't know exactly
> what
> >he did), and he says "it didn't work." He's very bright, though, so I
> trust
> >he went through reasonable efforts, but I thought I would check with you
> >all, since there's no point re-inventing the wheel.
>
> Derek Van Dyk
> Australian Proteome Analysis Facility
> Building F7B
> Macquarie University
> NSW 2109
> Australia
>
> http://www.proteome.org.au/
> Ph: 61-2-9850 6205
> Fax 61-2-9850 6200
>
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