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| 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. |
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#1
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| 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! |
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#2
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| 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? |
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| antibodyhelp , antiglurr1 , glur1 , glur1. millipore , problems |
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