homemade prestained proteins...

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Thu Sep 23 05:35:11 EST 2004

peter willingmann wrote:

> Hi Netters,
> i would like to make homemade prestained protreins for SDS-PAGE. I have 
> found a method wich used Remazol Dye and 50 mM Tris/038M Glycine 
> described from T.S. Piallay.. In my hand its not working properly
> e.g. the stain is not intense or the labelling efficency of the dye to 
> the proteins is not effective... I was using BSA, Ovalbumin, Lysozyme etc..
> Do anybody have a better method or could give me a tip...!!??

SH-containing proteins can be very efficiently labeled with IAEDANS, a
fluorescent dye. The NH-reactive fluorescamine has also been suggested. 

But I prefere multi-coloured standards, which are too laborious to
prepare in the lab. Also the MW of standards changes depending on the
number of dye molecules bound, manufacturers certify the apparent MW for
each batch of markers. Thus, everything considered, you are probably
better off by just buying the stuff. 

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