Is it possible to perform an electrophoresis to resolve different GAGs, using the standard acrylamide/bisacrylamide gels? If not, what would be the best way to do this? We would like to test it using non-fractionated heparin. Would it be possible to evaluate the different molecular weights of the different heparins in this solution?
Electrophoresis on cellulose membrane may be a solution for separating according to charge density not with the molecular mass
Follow the suggestion by Fabio
Also Sven Bjorson Has published excellent separation of gags using agarose gels
Capillary electrophoresis is another alternative but you need the instrument
In my opinion, PAGE of GAGs works quite well in general. I would suggest the boric acid buffer system (see protocols from Robert Linhardt). However, resolving unfractionated Heparin is quite tricky. Strong anion exchange HPLC is in my opinion the method of choice to separate complex mixtures of GAGs.
A standard PAGE is usefull to assess the molecular weight of GAGs, but if you want to resolve differently sulphated GAGs you should use 1,3-diaminopropane acetate agarose electrophoresis or cellulose acetate.
- for diaminopropane agarose electrophoresis see the work from professors Mauro Pavão and Paulo Mourão
- for cellulose acetate see the work by professor Nicola Volpi
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