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At 12:58 PM +0000 2/2/06, Paul McEwan wrote:



I'm currently working with a glycoprotein and am having great difficulty deglycosylating it for crystallisation. I can completely remove the sugars very easily when the protein is denatured (not much good when you're wanting to crystalise it), but seems completely resistant to treatment with endo H or pngase F when done under native conditions. Any hints on getting this to work would be greatly appreciated.

Perhaps you could try a sequential exo-glycosidase(s) then endo-glycosidase treatment in order give the endoglycosidase better access?

- John

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