Hi,

I was doing protein purification today and i observed that my niNTA beads turned brown after i bound my lysate with the beads. I eluted my protein but still the color of the beads were same.

I knew this situation occurs when DTT is added since DTT reduces the nickel.

But while performing two different protein purification I faced this problem in one. Although i used the same buffer composition for both the proteins.

Why it happened in one column not in case of the other protein?

My lysis buffer composition for both the proteins was

Tris,Back,1mmDTT,pmsf,BHCL,lysosyme,glycerol,10mm immidazole

Thank you.

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