We have also heard that a modest voltage (~ 50V) during swelling is beneficial but we haven't directly compared yet. Rarely do we get any proteins over 100 kDa. Apparently this is a physical limitation of the first dimension strips, I don't know if BioRad has overcome this.
Whoever can supply first dimension strips that can handle the larger molecular weights and a pH range beyond 10 (say up to 12 for nuclear proteins) would capture a large segment of the market.
--------------------------------------------------------
Gordon Alton, PhD
Gene and Protein Discovery Group
Signal Pharmaceuticals Inc.
5555 Oberlin Drive
San Diego, CA 92121
Email: galton@signalpharm.com
Phone: 619-558-7500 x8252
Fax: 619-623-0870
WWW: http://www.electriciti.com/signal/
-------------------------------------------------
-----Original Message-----
From: Katheryn Resing [SMTP:Katheryn.Resing@Colorado.EDU]
Sent: Wednesday, August 11, 1999 4:46 AM
To: Recipients of ABRF List
Subject: 2Ds and hi MW
What has been the experience of the IPGphore users out there with the
method of applying current while swelling the strips to enhance high
molecular weight proteins?
Can you do this with the BioRad system, and if so, are there any users out
there who have tried it?
Does anyone else have any tricks that enhance high mass detection. We're
getting proteins in 100-120 kDa range, but I know bigger ones are there,
because we see them on the 1D gel.
Katheryn