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Re: [ccp4bb]: glycoprotein
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Hi,
Another enzyme to try is PNGaseF, also from Hampton.
I have tried both EndoF1 and PNGaseF, both seperately and in combination,
and the PNGaseF alone is probably the best result wrt homogeneity on a gel.
The purification method uses an affinity step which means the
deglycosylases go straight through the column.
As yet though we have no crystals, but I am still trying.
Hope this helps
Louisa
PS if you want I could email a pic of the gel so you can see!!
At 12:44 PM 1/28/02 +0900, Kyung Hyun Kim wrote:
>Hi,
>
>This is not directly related to CCP4 packages. I have been trying to
>purify and crystallize a protein which is highly glycosylated.
>Deglycosylation using chemical and enzymatic methods is followed by many
>different chromatographic procedures. But, no crystallization luck yet.
>
>I wonder how people out there approach for the crystallization of these
>glycosylated proteins. Particularly, any new method for deglycosylation
>and purification would be highly appreciated. Thanks.
>
>K.H.