Go Back   Molecular Biology Forum Life Science Forums > Molecular Research Topics Forum > Protocols and Methods Forum
Register FAQ Members List Calendar Science Groups New! Arcade Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Stripping Western blots

Stripping Western blots - Protocols and Methods Forum

Stripping Western blots - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Reply
 
LinkBack Thread Tools Display Modes
  #1 (permalink)  
Old 10-26-2007, 06:31 PM
Ann Sutherland
Guest
 
Posts: n/a
Ann Sutherland RSS Feed
Default Stripping Western blots



Hi,
You're right - better to describe the already tested methods. The
methods we have tried are:

1. 1% SDS in PBS overnight at 37degreesC. This has variable results
depending on the antibody, but doesn't result in much loss of protein.

2. 2% SDS, 100mM beta-mercaptoethanol, 62.5mM Tris pH 7.4, for 30
minutes either at 50 degreesC or at room temperature. This works for
blots of cultured cell lysates, but results in high loss of protein
so it is too harsh for blots of embryo protein.

We're looking at the degree of phophorylation of several proteins, so
we need to have good removal of antibody, but we have very little
protein, so we need to minimize protein loss.
Thanks,
Ann

Reply With Quote


Reply

Tags
blots , stripping , 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
Stripping Blots Protocols moleculardude Western Blot Forum 3 10-31-2007 08:00 PM
Stripping western blots Ann Sutherland Protocols and Methods Forum 2 10-26-2007 01:36 PM
Normalization protein for Western blots? Peter Frank Protocols and Methods Forum 5 02-21-2007 11:43 PM
Special treatment required for dried Western blots? paul_wary@yahoo.com Protocols and Methods Forum 4 05-18-2006 11:06 AM
stripping antibodies from western blots Carl Ziess Protocols and Methods Forum 1 09-26-2004 05:31 AM


All times are GMT. The time now is 06:43 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2009, Jelsoft Enterprises Ltd.
Copyright 2005-2009 Molecular Station | All Rights Reserved