Hello, the issue I am facing is I would like to put a his tag on the N-terminal of my GPCR, but it has a signal peptide for the first 20 or so amino acids and I am unsure of how to get around this issue. Any help would be greatly appreciated thank you.
Either put the His-tag on the C-terminus, or arrange the construct so that the signal peptide is at the N-terminus and the His-tag is next to it.
Mark the position of cleavage of your signal peptide. As Adam said put His tag just after the cleavage site. So, after the cleavage you His tag will be there!
Carefully design your construct with his-tag after ss. From my experience His-tag on the C-terminus is better for protein with ss because your protein is seven-transmembrane domain receptors with N-terminus at extracellular site. What is the purpose of poly-histidine tagging ?
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