molar ratio of NTA and His-tagged protein

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Aggi
Aggi's picture
molar ratio of NTA and His-tagged protein

Hi all,

I'm trying to conjugate a his-tagged peptide to a molecule containing NTA. I was wondering whether there is an ideal molar ratio for each so that the conjugation will work. Also the concentration of the Ni2+ in solution? Maybe also other requirements such as oH and salt concentraitons in the buffers?

thank you

Anggia

R Bishop
R Bishop's picture
Interesting. Ive been

Interesting. Ive been reading up on NTA-crosslinking a bit today. Here's a reference that may be of use to you.

A self-assembled monolayer for the binding and study of histidine-tagged proteins by surface plasmon resonance.Anal Chem. 1996 Feb 1;68(3):490-7.
Sigal GB, Bamdad C, Barberis A, Strominger J, Whitesides GM.

pH 7.2 seems best. Still looking for the amount of nickel to add. Any luck on your end?

Rb