Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Molecular Biology Techniques
Register Search Today's Posts Mark Forums Read

Molecular Biology Techniques Molecular Biology Forum. Includes forums for common molecular biology techniques.


Totally blank western membranes!

Totally blank western membranes! - Molecular Biology Techniques

Totally blank western membranes! - Molecular Biology Forum. Includes forums for common molecular biology techniques.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 04-10-2012, 03:08 PM
Pipette Filler
Points: 2, Level: 1 Points: 2, Level: 1 Points: 2, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Apr 2012
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Unhappy Totally blank western membranes!



Hi there,

Apologies if this has been asked a few times before but it's a recurrent problem with me and seems to rear it's ugly head a few times a year.

Blank membranes - I mean completely blank - no MWM or anything!

I normally load between 50-100ug of protein, run at 40mA until loading dye runs off gel, transfer overnight, ponceau (which incidentally always shows bands), wash off with dH20 quickly then into PBS-T, block for 1 hour with 5%milk powder in PBS-T, primary also in 5%milk (either overnight or 1 hour at RT), wash x 3 with PBS-T, then into secondary for 1 hour before washing x 3 with PBS-T and developing with ECL.

There seems to be no rhyme nor reason for this - ie one day, I get bands and another day nothing!

Anyone shed any light?
Reply With Quote
Reply

Tags
blank , membranes , totally , western


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
Blocking Membranes for Western Blotting darkwng Western Blot Forum 5 07-08-2009 03:55 PM
Recommended Western Blot Membrane Supplier labratty Western Blot Forum 4 01-24-2009 01:40 PM
Storage of PVDF membranes after western cintiadp Western Blot Forum 7 10-10-2007 04:05 PM
How to Store Western Blot Membranes Later Use pipette_man Western Blot Forum 1 08-27-2007 04:31 PM
Silver staining Western membranes EK Protocols and Methods Forum 14 09-27-2003 02:37 PM


All times are GMT. The time now is 04:15 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.11809 seconds with 16 queries