Protein purification with sephadex? - (Sep/07/2006 )
I hope someone can help me. I want to do high throughput protein purification in a 96 well format (His tagged protein). Once I elute my protein from the NiNTA plate, I will need to do an imidazole removal step. I was thinking sephadex. My problem is that I am not sure which size/type of sephadex is best for this application. Would it have to be specific to each protein (assuming I am purifying from many different sources)?
ANY advice would be GREATLY appreciated!
you can desalt (buffer exchange) with sephadex g-10. this can be done with small columns or with spin columns (volume dependent). just follow the instructions.
PD-10 columns are very simple to use with a dilution factor of ~2.5 after desalting. Those are prepacked G25. You can get them from GE/Amersham/Pharmacia.