1 reply to this topic

[biobio]

Hi there!


I have recently made a His-tagged version of my gene of interest (a potassium channel) which I plan to overexpress in vivo in dorsal root ganglia neurons via a lentivirus. The idea is to try to visualise localisation of the protein in cell bodies and axons by immunohistochemisry using an anti-His antibody. I know these work well with immunoprecipitation/FACS but I don't really have any experience with them. So I just wanted to ask whether anyone has successfully performed immunos for His in vitro or in vivo and if yes what is your antibody of choice. And any other advice would be most welcome!

- Chris

[BioMiha]

biobio, on Nov 15 2009, 01:15 PM, said:

Hi there!


I have recently made a His-tagged version of my gene of interest (a potassium channel) which I plan to overexpress in vivo in dorsal root ganglia neurons via a lentivirus. The idea is to try to visualise localisation of the protein in cell bodies and axons by immunohistochemisry using an anti-His antibody. I know these work well with immunoprecipitation/FACS but I don't really have any experience with them. So I just wanted to ask whether anyone has successfully performed immunos for His in vitro or in vivo and if yes what is your antibody of choice. And any other advice would be most welcome!

- Chris

A friend of mine has done lots of experiments (=ELISA, WB, IHC) with a His-tagged scFv and anti-His antibodies. It works with no problems. I think she ordered mouse anti-His from Sigma, so she could detect the mouse antibodies with goat anti-mouse-HRP conjugated secondary antibodies (Jackson Immunoresearch). In our experience the antibodies from Jackson are the best you can get, but I don't think they actually sell anti-His antibodies.
Hope it helps.
Miha