d (on behalf of Wei Li
>)
>Subject: [ccp4bb] FW: [ccp4bb] highly glycosylated protein
>To: CCP4BB@JISCMAIL.AC.UK
>
>
>
> Dear Pascal and Matthias,
>
> I am sorry for the delay of reply, thanks very much
> for your suggestions on the glycosylation protein.
>
;
> BR
>
>
>
> From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Wei Li
> Sent: Tuesday, May 17, 2011 9:36 AM
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: [ccp4bb] FW: [ccp4bb] highly glycosylated protein
>
>
>
>
>
> Dear Pascal an
– surprised the cells live at all.
BR
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Wei Li
Sent: Tuesday, May 17, 2011 9:36 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] FW: [ccp4bb] highly glycosylated protein
Dear Pascal and Matthias,
I am sorry for the delay of
: [ccp4bb] highly glycosylated protein
Try to get hold of GntI deficient HEK293S cells (not commercially available).
Expression takes two weeks but you can achieve comparable yields to HEK293T.
These cells yield very homogenous bands on SDS PAGE. However, check also for O
glycosylation prediction.
As
Hi Wei,
Glycosylation usually stabilize proteins although it is a source of
structural heterogeneity for us crystallographers.Since you are expressing
in HEK293 cells, there is a strain of cells that is deficient for
glycosylation (it was designed by Gobind Khorana at the MIT I believe). You
may w
Dear all,
I am working on a high glycosylated protein, which was produced from HEK293
cells. Last time I added glycosylation inhibitor, the protein could exist some
monomers and dimers in a buffer pH5.4. This time I didn't add glycosylation
inhibitor, the protein seems highly aggregated and