Dear all
I tried a protein purification with L-Arginine 0,4M but my protein doesnt bind in HisTrap, then i reduced it to 0,2M and in the rest of the buffers I used a 0,4M concentration. With this protocol my protein doesn't stabilize in one of the peaks so if anyone could help me with a effective protocol to stabilize the protein with L-arginine or other additive.
Thank you
Hi there,
Arginine is not suitable for nickel affinity purification as it interferes with the matrix. To stabilize your protein you might play with pH (nature of the buffering agent), ionic strength and additives (like glycerol).
Ok. Do you know other additives that can a better stabilization than glycerol?. Thank you
David, keep in mind you can lose some stabilizing cofactor in gel exclusion chromatography which could explain loss of stability in one peak compared to the other... ;-)
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