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Glyoxime

Member Since 18 Feb 2005
Offline Last Active Dec 26 2012 09:31 AM

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Topics I've Started

E-book on recombinant protein expression

20 December 2012 - 11:37 AM

Hi everyone:

A Research topic and e-book about "Recombinant protein expression in microbial systems" is in the works. They're welcoming any type of work on the field.

More info:

http://www.frontiers...expression/1381

Disc SDS PAGE on Multiphor II system

07 December 2012 - 01:05 PM

Hi,

We'be trying to run discontinuous SDS-PAGE gels on a semi-dry flatbed system (Multiphor II from GE). As proposed in the book Electrophoresis in Practice, we used:

Cathodal and anodal Buffer: Laemmli Buffer 5X
Stacking Gel: Tris-HCl 1.25M pH 6.7 / 0.1% SDS
Resolving gel: 12% homogeneous, Tris pH 8.8 like a vertical gel
Power settings: 600V, 50 mA, 22 W, 15 C

It was a complete mess, it took more than 8 h to run. Several bands turned yellow and stopped running down the gel. We tried lowering Tris to 0.125M like a normal vertical gel but the samples were not resolved at all (the MW standard looked OKish).

As everyone, we use the vertical format system for SDS-PAGE, but we were trying to do SDS-PAGE in the Multiphor so that we would have enough practice before doing 2D gels.

Has anyone tried this protocol?

I hope 2D gels are easier...!