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Smear at sides of a lane.


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#1 Al Bunsy

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Posted 28 January 2005 - 09:50 AM

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!

#2 thefallguy

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Posted 29 January 2005 - 05:22 PM

Al -

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

#3 Al Bunsy

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Posted 30 January 2005 - 06:47 AM

>> 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...

:angry:

#4 thefallguy

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Posted 30 January 2005 - 07:57 AM

>>  thefallguy

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

:angry:

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

#5 Al Bunsy

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Posted 30 January 2005 - 09:43 AM

Come on!

Just to kill someone. It helps.




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