Hi all,
We are looking for an antibody that recognises N-terminally tagged His tag on a recombinant protein, any recommendations or where to find this information would be appreciated.
Thank you
Most His-tag antibodies are designed to recognize the His-tag sequence regardless of its position (N-terminal, C-terminal, or internal) in the protein. This is because the His-tag sequence itself is what the antibody binds to, not its position within the protein.
You can try His Tag Monoclonal Antibody (6G2A9) by GenScript or His-Tag Antibody by Cell Signaling Technology. However, some antibodies might have a preference for N- or C-terminal tags due to steric hindrance or other factors. For this you can try N-His tag (PODI-0077) by ImmunoPrecise
To find an antibody that recognizes an N-terminally tagged His tag on a recombinant protein, you can follow these steps:
By following these steps and conducting thorough research, you should be able to find an antibody that recognizes N-terminally tagged His tags on recombinant proteins.
Sigma Santa Cruz etc. etc. so many antibodies are there which are specific to anti histidine
It would be interesting to know why you need an antibody specific for N-terminal His-tag. Usually, you don't need to check where the tag is located, as it is added by you, so you should know in advance where it is. Some antibodies might have slight preference for one or the other location (depending on the protein structure) but usually His-tag antibodies do not bind to the tag on both ends differently. In fact, His-tag antibodies are not the easiest one to generate with good affinity, let alone specificity for one terminal position.
Hi all, thanks so much for your answers. We have produced a C-terminal specific anti-His Tag antibody in plants, and even saw it binds only to His Tags on the C terminus. I am not sure exactly why- would be good to understand. Someone was asking us for one that recognises N terminal His tags as that is the location of their His Tag, hence my question.
Hi, maybe you should confirm that it binds only the His-tag and not one or more of the upstream amino acids. Alternatively it could have a specificity towards the carboxy-group of the last His which could explain its preference.