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Deglycosylation problem

Deglycosylation problem - Protein Science

Deglycosylation problem - Post Questions and Discuss Protein Structure and Function, Protein Localization etc.. Please do not post Proteomic questions here, there is a Proteomic Forum below.


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Old 09-10-2009, 08:52 PM
Pipette Filler
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Default Deglycosylation problem



Hi
I'm working with glycoprotein and have 2 glycosylation sites. i mutated one and other one I'm trying to get rid by digesting it with Endo H. I did small scale and it worked fine but when I did large scale i still see not fully deglycosylated band on my gel. Somehow the the protein is not getting fully deglycosylated, i tried using 10 fold more enzyme + more time still no change. How can I fully deglycosylate my protein since little bit of heterogenity will interfere with my crystal structure. Plz let me know!

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