Hello,

I am working with a G-protein coupled receptor. To be honest, I am not sure of the glycosylation of the protein. It turns out that transient transfections followed by western blotting gives a large molecular weight smear. There doesn't appear to be any problem with the antibody. Have there been reports of glycosylation causing this? If so, what is to be done. All I really want right now, is a clear expression band for my protein.

Thanks

There are several faqs on this site re: glycosylation. The simplest thing is to treat your recombinant protein with PNGaseF enzyme to attempt to remove N-glycosylation, then run the gel. Get back to us if that doesnt work and we can discuss ways to remove O-glycosylation.

Places to look on SS
in the post-tranlational modification forum in Protein Chemistry or assays in Protein Chemistry.

Good luck

Rb