I need an easy way to immobilize a polysaccharide (xylan) on a cover slip glass surface for one-molecule analysis using confocal fluorescence microscopy.
Hi Gustavo, does you polysaccharide have any other functional groups besides -OH? Does it have an aldehyde group?
Hi, Steingrimur. Thanks for your answer.
The polysaccharide I am talking about is the xylan, so it supposedly has an reducing end, whose open-chain form presents an aldehyde group.
Hi Gustavo, since xylan (hopefully) has an aldehyde group, you can try coating your glass surface with an aminopropyl silane
http://www.sigmaaldrich.com/catalog/product/aldrich/440140?lang=en®ion=US
You should acid wash the glass surface with H2SO4 at RT, preferably ON, then wash the acid away with DiH2O, incubate the glass with 10% aminosilane in 100% ethanol, or 100% methanol. (Avoid alcohols containing H2O, because this can cause aggregation of the silanes.
After 1 hr incubation remove the glass and let it dry for ~30 min at RT. Then add your xylan in buffer that does not contain a primary amine to the glass and let it dry. After ~30 min wash away salt residue.
If the xylan has an aldehyde residue, then (hopefully) some of them will be attached to the aminosilane through Schiff base.
Let me know how things work out.
Hi, Steingrimur.
I'll try to follow your suggestion and post here the results of my attempts.
Thank you for your help.
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