Electrophoresis problem

Andrew Warkman awarkman at julian.uwo.ca
Wed Mar 24 07:54:07 EST 1999


> Do you mean stain the gel in coomassie, destain, and then electrophorese
> the protein out of this same gel onto the nitrocellulose?  That wouldn't
> work because the SDS would be leached during the staining and destaining
> procedure, and the transfer of the proteins would be minimal at best.
> The transfer would also be in both directions, and you would have to
> place nitrocellulose on both sides of the gel when you did the
> electrophoresis.

Not true..

(1) if SDS is in the sample loading buffer (and assuming that the samples
are boiled before loading) the SDS will be permanently bound to the proteins

(2) SDS bound to the protein serve to masks the native charge and gives the
entire protein a negative charge (with the same chare to mass ratio) thus
all protein would be electrophoresed (I prefer the term  electroblotted)
toward the positive electrode

Cheers

Andy


**************************************************
Andrew Warkman
The University of Western Ontario
Department of Zoology (B&G 344)
London, Ontario, Canada
N6A 5B7

http://publish.uwo.ca/~awarkman/
**************************************************





More information about the Methods mailing list