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