I am looking for the antibody or reagent that allows the imaging (immunostaining or western blot) of internal 10x his tag. I tested several his-tag antibodies to see them by western blot but failed. Is there any recommended antibody or imaging reagent for 10x his tag?
Dear Ah-Ram Kim see link below may be useful for you.
Good luck!
https://www.labome.com/method/Histidine-Tag-and-Anti-Histidine-Antibodies.html
I had good luck with Nickel-HRP, as an antibody-like detection reagent for his-tags in Western.
Hi Ah-Ram,
- Have you tried to use the anti-his antibodies with a positive control (well known his-tagged protein) to see if you could get a result?
-You may need to strip your membrane to make his-tag more accessible to the antibody. Sometimes, even though the protein is denatured by loading buffer and heat, a tag or epitope(s) aren't accessible.
-Increase the first Ab concentration, incubation time, and the exposure time .
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