I have isolated various marine polysaccharides (fucoidans) from several seaweeds and would like a rough estimation of their sizes (kDa) by running them on a carbohydrate polyacrylamide gel electrophoresis (C-PAGE). However, I cannot find a carbohydrate molecular marker/ladder to run it against.
There are oligosaccharide standards available. For instance, I have chitosan oligosaccharides from YSK (http://www.yskf.jp/yskf_en/yskf_en_03.html) which are 3 - 7 monomer units. If you want larger sizes you may find what you want if you look for HPLC standards. Or carry out your PACE experiment, cut out your bands, and measure the size of your isolated carbohydrate by MALDI-TOF or some similar MS technique which can measure large masses.
(MALDI-TOF can measure up to 30'000 m/z, which is one 30kDa molecule with a single charge.)
Thanks Sally!
I am doing GC-MS and MALDI-TOF for in depth chemical analysis of my compounds. However, up until this point I have not been succesful. On MALDI i've tried various matrices and columns but the polysccharides are too heavily sulpahed and negatively charged. I've hydrolysed them too and tried MS techniques to no avail. The C-PAGE was just for a rough estimation before sending the samples off for MS again. I've done SEC-HPLC which gave me a rough estimation of size too. The one polysaccahride was >2000 kDa.
I will look out for the standards you suggested. That page you suggested is not in English though.
I appreciate the insight.
Regards,
Grant.
Sorry. It's a Japanese company but the page does have an English version. Look in the top right hand corner. Then click on Functional Materials World in the next page.
I actually linked to an English page, but you seem to have got diverted to the site map in Japanese. Still, if it's not chitosan oligosaccharides you're interested in you needn't waste too much time there.
By the way, 2000 kDa would be about 11 monomer units.
Found what i was looking for. Anyone else interested in oligosaccharide molecular ladders check out:
http://www.lifecore.com/research-products/oligos-mw-ladders
Actually speaking you do not get molecular markers specifically made for C-PAGE. However, you can make your own set of markers, using the primary standards of HPLC or HPTLC, but then you must decide what to mix and what not. It is a trial and error method, and may consume some of your precious time for a particular condions of C-PAGE.
Thanks Jai. I was thinking of making my own too but you are correct, optimization will take some time.
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