anti his antibody for western blotting
Posted 03 November 2010 - 12:02 AM
I am doing some western blotting for an intracellular his tagged recombinant protein in CHO cell lysate, I tried anti his HRP from roche and anti penta his from qiagen followed by conjugated second antibody. with anti his HRP I didnt get any band even in 200 microgram lysate but giagen antibody in its minimum titer (1/1000) has high background. does any body has any suggestion for an anti his antibody with high sensitivity and specifity in crude cell lysate?
Posted 05 November 2010 - 02:14 AM
Posted 06 November 2010 - 04:38 AM
I used goat anti fab antibody (affinity purified from anti fab poly clonal antibody)from sigma, my coworkers are using 1/3000 titer for purified samples I tried 1/3000, 1/6000, 1/10000 titers and I always had a lot of non specific bands.
We've done some stuff with an anti-his mAb from Sigma. It seems to work OK. My question is what sort of conjugated secondary are you using (specificity, company).
Posted 06 November 2010 - 12:23 PM