4 replies to this topic

[Al Bunsy]

Hello dear all.

My non-denaturing PAGE looks a bit non-publisheble because of a smear at sides of lanes, so, my proteins are not like bands, rather like comets with two tails.

Sure, the problem might be easily solved, but I don't know how.

Can you give me some comment on my problem?

Just in case: 5% stacking gel, 12% separating gel, Tris-Glycin running buffer, 40-80 mkg of protein per lane (and I'm afraid I can't use less because it's in-gel enzyme activity experiments, there is not much emzyme in my samples).

Thank you!

[thefallguy]

Al -

the comet like tails are due to salt. Is dialysis an option at all for you or will the proteins precipitate?

[Al Bunsy]

>>  thefallguy

Thanks a lot for the comment.

My protein samples are dissolved in 1xPBS, I don't think there is a lot of salt in it...

Actually, I found some notations on problems like mine.
It might be useful for everybody, so, I'm placing the link here (hopefully, I'm not breaking rules of the forum):

http://www.mirador.c...b_05Mar2004.pdf
It's the troubleshooting guide for protein electrophoresis.

Good luck to all of us, soldiers of desperate fight against the Evil of Darkness. Heh...

[thefallguy]

Al Bunsy, on Jan 30 2005, 07:47 AM, said:

>>  thefallguy

.....................
Good luck to all of us, soldiers of desperate fight against the Evil of Darkness. Heh...

Soldiers of desperate fight against Evil of darkness, eh, Al? Looks like you badly need a vacation or a change of life or both

[Al Bunsy]

Come on!

Just to kill someone. It helps.