How to run poly-lysine on tris-tricine gel - (Apr/03/2007 )

Hey guys,

I would really appreciate any input I can have on this subject. I have been trying to figure out ways to run my short poly-lysine peptide on 10-20% Bio Rad tricine ready gel. The gel was run at 100V and I stained it with Coommassie. After staining all I could see was my peptide Mw ladder but never the poly-lysine: None at all. These poly-lysine samples range in size from 60-mer to 200-mer so their Mw's are from ~12 kDa-40 kDa.

I wonder if this had anything to do with the fact that poly-lysine is highly basic at neutral pH? Has anyone ever run polylysine on a gel?

Thanks so much for your input!

-greentea-

Yes, they carry positive charges and go to negative electrode. Swtch to acetic acid or Mops buffer with a lower pH (pH 4-5) and flip the polarity of your apparatus, you will see they move into the gel. You cant use your standards, though. They are not designed to perform at that way.

-genehunter-1-

QUOTE (genehunter-1 @ Apr 3 2007, 06:16 PM)

Thank you very much for your reply! I am new to this business and I greatly appreciate any help I can get

Would you mind telling me the compositions of the acetic acid and/or MOPS buffer? Thanks.

-greentea-

50 mM of HOAC or MOPS, pH adjusted to 4.0 to 5.0 with NaOH/HCl will do the job.

-genehunter-1-

QUOTE (genehunter-1 @ Apr 3 2007, 08:02 PM)

50 mM of HOAC or MOPS, pH adjusted to 4.0 to 5.0 with NaOH/HCl will do the job.

Great! I will try that. Thank you very much!!!

-greentea-