Has anyone used Millipore's polyclonal GluR1 antibody to do a western blot? We recently purchased some and even at 1:500 dilution I could not get any bands- other antibodies run simultaneously worked fine. I have tried 1:4000, 1:2000, 1:1000, and 1:500 dilutions. I keep coming up with very faint smudge of signal, but no specific bands.
If anyone has used this antibody and gotten it to work, I would really appreciate it if you would tell me what you did to get results with it.
I have been dissolving it in TBST like I do with all my antibodies, inclubating for 1 hour in primary AB and 2 hours in secondary AB with 3 10-min washes in TBST between the ABs and after the secondary, and finishing with 1 minute in ECL reagent then exposing for 2-3 minutes. I can't get any good results!