REFOLDING - visualisation of protein after denaturation step? - (Dec/20/2006 )
Just checking here,
after the initial denaturation with either Urea or GnHCL, then after the spin to bring down the debris, your protein should be in the supernatent, right? I mean, if it is not there then it will stil be in the Inclusion bodies, its not as if when you refold the protein is going to appear?
cheers
-grahamkeith-
QUOTE (grahamkeith @ Dec 20 2006, 05:37 PM)
Just checking here,
after the initial denaturation with either Urea or GnHCL, then after the spin to bring down the debris, your protein should be in the supernatent, right? I mean, if it is not there then it will stil be in the Inclusion bodies, its not as if when you refold the protein is going to appear?
cheers
after the initial denaturation with either Urea or GnHCL, then after the spin to bring down the debris, your protein should be in the supernatent, right? I mean, if it is not there then it will stil be in the Inclusion bodies, its not as if when you refold the protein is going to appear?
cheers
yes if your protein is denatured, after the denaturation step it should be in the superatant and visible on SDS-PAGE.
High concentration of GnHCl can messed up the gel, so if you want to analyze this supernatant,
you should load the sample on the side of the gel.
Sophie
-Sosous-