|Register||Search||Today's Posts||Mark Forums Read|
|Western Blot Forum Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.|
| ||LinkBack||Thread Tools||Display Modes|
Problems with GLUR1 antibody-HELP!
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!
Re: Problems with GLUR1 antibody-HELP!
I've been using Millipore's GluR1 ab for months w/ excellent results UNTIL using a new vial from a shipment received this month (April '09): lot# LV1563305 (cat#1504). I got absolutely no signal (usually get a clear, clean band at 105kDa, after a short exposure). Next, tried the 2nd vial of antiGluR1 from the said shipment/same lot#, and this time in parallwl w/ a colleague's aliquot of AB1504 from an older lot#. The older/different lot# ab worked perfretly, as usual, and again lot# LV1563305 gave absolutely nothing! Currently trying to get the vender to send me 2 new vials from a different lot#... is this ur lot number?
|antibodyhelp , antiglurr1 , glur1 , glur1. millipore , problems|
|Thread||Thread Starter||Forum||Replies||Last Post|
|Problems in screening genomic DNA Library with antibody||srivels||Molecular Cloning Forum||0||01-17-2010 03:42 PM|
|Precipitated Antibody Problems||Biomencer||Antibody Forum||1||06-23-2009 09:17 AM|
|Production of Monoclonal Antibodies - Technique in Mouse||Cellbiogal||Antibody Forum||2||01-08-2009 09:52 PM|
|Antibody Problems||patingsadagat||Protocols and Methods Forum||5||01-09-2008 06:44 AM|