Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protein Science > Western Blot Forum
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.


Problems with GLUR1 antibody-HELP!

Problems with GLUR1 antibody-HELP! - Western Blot Forum

Problems with GLUR1 antibody-HELP! - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 04-09-2009, 04:34 PM
Pipette Filler
Points: 65, Level: 1 Points: 65, Level: 1 Points: 65, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Apr 2009
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default 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!
Reply With Quote
  #2  
Old 04-16-2009, 01:47 PM
meb meb is offline
Pipette Filler
Points: 2, Level: 1 Points: 2, Level: 1 Points: 2, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Apr 2009
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default 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?
Reply With Quote
Reply

Tags
antibodyhelp , antiglurr1 , glur1 , glur1. millipore , problems


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Problems in screening genomic DNA Library with antibody srivels Molecular Cloning Forum 0 01-17-2010 04:42 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 10:52 PM
Antibody Problems patingsadagat Protocols and Methods Forum 5 01-09-2008 07:44 AM


All times are GMT. The time now is 08:29 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12228 seconds with 16 queries