Dear Pascal and Toufic,
Many thanks to both of you for the pointers. Toufic, I actually poked
around online and bought exactly the same kit that you are suggesting,
especially because the beads are reusable. So your experience is
reassuring. So I'll find out soon.
I also plan to play around with
Hi Raji,
I addition to the tips from Pascal, I would like to say that for a memb
protein I worked on with a his-tag separated by a thrombin site, I used
thrombin cross linked to agarose from Sigma (1 mL). The beads can be
collected, washed and reequilibrated, making it ready for use so many times.
Hi Raji,
Thrombin is a rather good protease and behaves well in a large set of
different detergents ( there is a paper by Michael Wiener that describes
the relative efficiencies of several usual proteases, amongst those
thrombin is inlcuded, used routinely for cleavage of membrane protein
fusions
Hi Folks,
Sorry this isn't a non-ccp4 post.
I am working with a membrane protein for which I am finally able to scale
up expression. I am now also able to partially purify my protein from a
medium-scale (12-18L) bacterial culture using a two-step tandem affinity
purification protocol (Talon follo
