I have it on good advice from a 2D gel expert, Ben Herbert at the
Australian Proteome Analysis Facility, that given favourable circumstances,
it is possible to load up to 30 mg of protein by rehydrating the
immobilized pH gradient strip in the sample, a method he devolped some
years ago.
Every time I see Ben's gels of wool proteins (go figure the solubility of
those !) with such massive amounts of protein on a single polyacrylamide
gel I am gobsmacked (to use a British expression).
Ben can be contacted on bherbert@rna.bio.mq.edu.au
Regards...Ken
>Date: Fri, 16 Jan 1998 08:53:24 +0100
>From: Amanda Hall <mbah@medicine.newcastle.edu.au>
>Subject: re:IEf gels or strips
>Old-To: abrf@aecom.yu.edu
>MIME-version: 1.0
>To: Recipients of ABRF List <abrf@aecom.yu.edu>
>Sender: Association of Biomolecular Resource Facilities
><abrf-request@aecom.yu.edu>
>Precedence: bulk
>
>Dear David,
>
>I am only a beginner with 2D gels but in some ways I think that this makes
>me more qualified to comment on at least the ease of use of immobilised
>strips.
>
>I am currently trying to get this procedure up and running and have had
>experience with both tube gels and immobilised strips. I am currently using
>the Pharmacia Immobiline system of Immobilised strips and have found it
>very easy to use and run. The strips are especially easy to transfer from
>the 1st dimension tothe second dimension run. Pharmacia ahve a new
>reswelling cassette where you can rehydrate the strips with the sample so
>that sample application and rehydration are completed in 1 step. This
>removesthe need for sample cups or any thing making the procedure even
>easier. You are supposed to be able to reswell the strip with up to 5 mg of
>protein. I have not tried a sample this high. The strips are covered with a
>mineral oil for the run so the chance of contaminants from the system is
>fairly low.
>
>i hope that my somewhat limited and probably naive opinion has been of
>assistance
>
>Good luck
>
>
>Amanda HAll
>
>
>Amanda Hall
>Professional Officer
>Newcastle Protein
>University of Newcastle
>
>ph 02 4921 7299
>email: mbah@medicine.newcastle.edu.au
>
********************************
Ken I. Mitchelhill
The John Holt Protein Structure Laboratory
St. Vincent's Institute of Medical Research
41 Victoria Parade
Fitzroy 3065 Victoria
AUSTRALIA
Telephone: 61-3-9288 2480
Facsimile: 61-3-9416 2676
Email: k.mitchelhill@medicine.unimelb.edu.au
Laboratory: http://www.medstv.unimelb.edu.au/WWWDOCS/SVIMRdocs/JHPSL.html
ABRF: http://www.medstv.unimelb.edu.au/abrf.html
***********************************