I am expressing my protein in E.coli expression system and to purify it I am gonna use His tag. After this i am going to use a method to measure the binding affinity between this protein and some other protein. I wanted to ask from dear scientist if i have to remove this His tag for LSPR or any other binding assay?
The His tag system in E. coli has a cut-off site for enterokinase, you must follow the protocol to obtain only your protein of interest. Then you can perform your binding tests. The answer is affirmative, after overexpressing it and purifying it to homogeneity you can remove the label of six histidines.
It isn't necessary to remove the His tag, but it is advisable if there is any possibility that the tag could interfere with the binding interaction.
The interaction of the immobilized binding partner with your protein should not be influenced by the His tag. If this is not the case, it is possible to exploit the His tag for binding of further His tag-specific ligands to increase your signal and perhaps to prove the specificty of the observed interaction.
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