2D electrophoresis

Benoit Viard benoit at nibh.go.jp
Wed Jan 24 05:17:23 EST 1996


Hello everybody,

I am actually trying to perform 2D gels with S. cerevisiae crude extracts.
But the resolution of the spots is not that good and I would like to
stretch the region of the gel comprised between pH 6 and 8, and molecular
weight between 15000 and 50000.

I use NaOH 100 mM and phosphoric acid 10 mM.

The ampholyte concentrations are :
1.6% ampholyte 5/7
0.4% ampholyte 3/10 in both first dimension gel and first dimension sample
buffer.

0.8% ampholyte 5/7
0.2% ampholyte 3/10 in sample overlay buffer.

How can I modify these concentrations to enlarge the interesting zone ?

Thanking you in advance.

                        B. VIARD, Postdoctoral Researcher

                        National Institute of Bioscience and Human-Technology
                        Biochemical Engineering Laboratory
                        1-1, Higashi, Tsukuba
                        Ibaraki 305
                        JAPAN





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