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Hyaluronate electophoresis

PAGE electrophoresis polysaccharide hyaluronate uronic acid

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#1 EvilTwin

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Posted 21 February 2013 - 01:04 AM

Hi everyone!
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?

#2 mdfenko

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Posted 21 February 2013 - 06:14 AM

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 ...

Edited by mdfenko, 21 February 2013 - 06:15 AM.

talent does what it can
genius does what it must
i do what i get paid to do





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