We have expressed and purified the recombinant viral protein found in the insoluble fraction. Please suggest some methods or reagents to solubilize the protein.
I guess you want to refold the recombinant protein, in that case you can solubilize the pellet using urea or guanidine HCL, afterward, you need to slowly decrease the concentration of these molecules to induce refolding.
Dear Akanksha Vyas
protein solubilization as suggested from Omar Gonzalez-Ortega can be easy achieved using denaturing reagent as urea or guanidine Hcl, however in this way you will obtain the unfolded proteins and i'm note sure is suitable for your purpose. Protein refolding, with replacement of the denaturing agents with standard buffers in fast way (direct refolding) by dilution or using a slow approach (by dialysis) is something that you can try but is very complex process, that need a sort degree of luck.
Are you sure that you cannot produce the protein in the soluble form? Did you performed some trials (eg test low temperature induction, different lisis buffers, different e.coli strains?) since in my experience in many cases is more easy to solve the insolubility problem at the origin instead obtain result with refolding.
good luck
Manuele
Dear Akanksha Vyas
You could adding NaCl (100-250 mM) to the culture medium and/or modify agitation speed.
Good luck
Maura
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