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.

messy running gel western blot

messy running gel western blot - Molecular Biology Techniques

messy running gel western blot - Molecular Biology Forum. Includes forums for common molecular biology techniques.

LinkBack Thread Tools Display Modes
Old 04-18-2013, 11:31 PM
Pipette Filler
Points: 57, Level: 1 Points: 57, Level: 1 Points: 57, Level: 1
Activity: 0% Activity: 0% Activity: 0%
Join Date: Apr 2013
Posts: 2
Thanks: 0
Thanked 0 Times in 0 Posts
Question messy running gel western blot

I am culturing SKNSH cells, 1)collecting the cells by trypsin and centrifugation, 2) washing the pellets with pbs.
Then I am adding RIPA buffer to lyse the cells by freeze-thawing. I add protease inhibitors and also DNAse to get rid of DNA precipitate. I am looking for a cell membrane protein.
My buffer recipe is as follows: 40mM TRis (pH 7.6), 120 mM NACL, 1% TritonX-100, .3% SDS & 0.5% Na-deoxycholate.
I add 4x laemlli buffer and boil at 95C for 5 mins. I run gel at 100V and make sure its not hot.
Despite all this my gel is running messy and even beta-actin band shows up broken & mess
Attached Images
File Type: jpg photo 4.jpg (88.6 KB, 0 views)
File Type: jpg photo 1.jpg (85.1 KB, 0 views)
Reply With Quote

blot , gel , messy , ripa buffer , running , viscous lysate messy gel , western , western blot

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
Western Blot Background Troubleshooting molecule2005 Western Blot Forum 18 10-07-2010 11:49 AM
Can we use dot blot inplace of Western blot? peterish Western Blot Forum 3 06-16-2010 07:59 PM
dot blot and western blot Liang Western Blot Forum 1 08-07-2009 03:26 PM
western blot question Rebecca Pickin Protocols and Methods Forum 2 07-25-2007 08:01 PM
Western blot problem -disappearing proteins! Jayakumar, R Protocols and Methods Forum 0 12-13-2004 02:21 PM

All times are GMT. The time now is 05:00 PM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2015, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.11275 seconds with 17 queries