Protein purification - (Jan/31/2006 )
Hi there. Any chance anyone could help with me with these pretty basic questions?
I need to remove all non-soluble proteins from my protein mixture... How would i go about doing this?
As well as, what is the basic protocol for HIS tag purification?
(And final silly question... what is the English word for the plastic "swiper" you use to "swipe" a colony off a plate? )
Thanks in advance!
Whats about centrifugation?
I use this step after lysation or frenchpress to seperate the crude into the supernatent and a pellet. You can try it in a centrifuge for tubes (Eppis) or especially in a "ultra"centifuge for higher g forces. 45 min at around 25000 g and there is no non-soluble protein in the supernatent.
to remove non soluble proteins, I would say centrifuge as fast as possible.
for the protocol for his tag protein purification have a look at the attached file.
there are also very good handbooks on quiagen website.
Do you mean inoculating or drigalski loop???