Hyaluronate electophoresis - (Feb/21/2013 )
I would like to ask if anyone has any experience in electrophoresis and detection of hyaluronate. I did PAGE of a digest mixture (consisting of HA chains of various lengths) according to numerous protocols (15% and 10% gel in TBE, linear gradient 20%-3% and 18%-1,5%, with samples in NaOAc, NH4HCO3, NaOAc+HCl, dH20
or TBE, with the voltage of 300V and 100V and detection by Alcian Blue in water, water+2% acetic acid or water+40%methanol+10% acetic acid.
Everytime I just got an ugly smudge instead of nice "ladder". Today I will try Alcian Blue+Silver staining protocol, but I doubt whether it can do any good. I think a smudge is a smudge, no matter how you dye it, though I still hope I am wrong.
Does anyone have any idea what else I could try?
you are correct, a smudge is a smudge.
you can try the procedure in this paper:
microanalysis of ...
it contains page and references to ce methods.
or you can see this paper for ce methods:
ce for analysis of ...