Albumin depletion from sheep plasma

[Pow Joshi]

On 04/03/2008, Arianmanesh, Mitra <r06ma6 from abdn.ac.uk> wrote:
>
>  Dear Pow
>
> Actually, I am doing Albumin depletion as I need the plasma for running
> the 2-D gel. if there is Albumin in the plasma, the gained 2-D gel would be
> very fussy and smeary due to lots of loaded Albumin as it covers other
> proteins on the 2-D gel. Now I'm trying Zoom fractionator for separating
> Albumin based on its PH, however it was not successful so far. I would be
> happy if you advise me.
>
> Sincerely,
>



I can only suggest that you do partial fractionation say starting with
ammmonium sulphate cut offs and then ion exchangers, if the Cibacron blue
does'nt work for you. These are some really ancient protocols but woork
well.
I understand the absolute frustration when there's this big blob of albumin
sitting messing up an entire gel run.....esp. if you're looking for really
low concentration species.

hope that helps
Pow

Mitra
> **
>
>  ------------------------------
> *From:* Pow Joshi [mailto:pow.joshi from gmail.com]
> *Sent:* 04 March 2008 00:05
> *To:* Arianmanesh, Mitra
> *Cc:* methods from magpie.bio.indiana.edu
> *Subject:* Re: Albumin depletion from sheep plasma
>
>
>
> On 03/03/2008, Arianmanesh, Mitra <r06ma6 from abdn.ac.uk> wrote:
> >
> > Dears
> >
> > I need to deplete Albumin from sheep plasma while there is no defined
> > and perfect method for it. Actually, I used several methods for that
> > such as Immunoprecipitation and SwellGel Blue Albumin Removal Kit but I
> > could not get good results. So, now I need to know that how to deplete
> > Abumin from the sheep plasma. Can anybody give me an advise to do that
>
>
>
> yes, that is a tough one.. Perhaps, you could just use Blue sepharose
> CL-6B (trade name I think is Cibacron), and run your plasma one it as a
> column a few times or perhaps do a serial culk incubation.
> Now, if you wish to purify antibodies, you could use the afffinity
> columns; if you don't care about having or not having antibodies in the
> plasma/serum, you could probably use an ion exchanger as well. ...actually,
> I think I am a bit fuzzy here, and can be able to help you better if you let
> us know what exactly you wish to get out of the plasma.
>
> best
> Pow
>
> well?
> >
> > Kind regards,
> >
> > Mitra
> >
> >
> >
> > The University of Aberdeen is a charity registered in Scotland, No
> > SC013683.
> >
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>
> The University of Aberdeen is a charity registered in Scotland, No
> SC013683.
>