Protein glycosylation - How to determine? (Jul/14/2009 )

I have isolated complete open reading frame of a gene from plant and now I want to express it recombinantly. For the proteins which undergo post-translational glycosylation, euokaryotic expression systems are suggested. In such case how do I know whether the protein that I am going to express in vitro is glycosylated in planta or not? There is no such information available about my protein of interest in the literature. If I isolate this protein from plant, is there any way know where it is glycosylated or not?

Yes -- you can use a general glycoprotein staining kit, like the Pro-Q Emerald 300 Glycoprotein Stain Kit from Invitrogen (see here).

If you isolate the protein from the plant, you can see if it's glycosylated in planta with this kit. Whether it will be glycosylated in another background is a question.

HomeBrew on Jul 14 2009, 07:26 PM said: