gel electrophoresis

[Waleed]

hello all.... i hve many questions and i wish that i can find answrs through this forum..:
1- whats the effect of PH in the buffer used for gel electrophoresis separtion.
2- why dont we use distilled water as a running buffer in the spearation through AGE.
3- if we changed the Voltage during separtion gradiently : 40. 50, 60 120 and 160 is that will give the same effeciency ?

[DR]

There are probably better forums than this one for electrophoresis questions and it's been 15 years since I've done any myself, but here goes (fwiw)

pH can be a very powerful tool for enhancing the separation of proteins via PAGE - google IEF and Phastgel for more details.

DI is highly resistive to electricity - no current=no migration of analytes through the gel

voltage changes (I think) would only make the separation take longer. You want to apply enough voltage to get things moving but not enough to cook the system. Lower voltages would probably just result in longer run times and more opportunity for band broadening via diffusion.