There are more than one way of refolding your protein, and it really depends on the nature of your protein. Mine is rich in cystine residues, after denaturing my protein in 6M GdmCl, I refold my protein using a method called rapid dilution where my protein solution with high conc of denaturant is diluted into large amount of solutions containing oxidised and reduced glutathione with vigorous stirring. Found this review online, hope it helps~

Hi there,

Dilution, dialysis or application of a refolding buffer on a column where the denatured protein of interest has been immobilized are the usual approaches.  

Thank you guys for your answer.

In the case of denaturation by urea, you will want to remove urea by either a desalting column or by dialysis into PBS. Hope this helps.

There are kits available with a set of mixtures that allow you to survey conditions for refolding. Google "protein refolding kit"

Thanks Adam

Important to be aware that there will always be a competition between a) the tendency for the protein to fold up correctly as evolution has taught it do and b) the tendency for partly-folded molecules that haven't yet managed to hide their hydrophobic patches to aggregate in an uncontrolled way and just precipitate out of solution. That means that you have to carefully control protein concentration (inevitably  high protein concentration increases the statistical chance of aggregation) and also that you need to experiment with buffers to control the balance between favouring hydrophobic vs. ionic interactions. People often assume a buffer is a buffer - i.e all it does is control pH. Not true. 

Thanks Paul