Go Back   Molecular Biology Forum > Molecular Research Topics Forum > Protein Science > Western Blot Forum
Register Blogs FAQ Members List Calendar Science Groups New! Arcade Search Today's Posts Mark Forums Read

Western Blot Forum Discuss western blotting and immunoblot in the western blot forum.

western Videos


Convex shape of sample bands on SDS PAGE gel

Western Blot Forum

Discuss western blotting and immunoblot in the western blot forum.



Register Molecular Biology Forums
Reply
 
LinkBack Thread Tools Display Modes
  #1 (permalink)  
Old 04-20-2007, 09:43 AM
Pipette Filler
Points: 733, Level: 14Points: 733, Level: 14Points: 733, Level: 14
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Apr 2007
Posts: 2
ssacket1 RSS Feed
Default Convex shape of sample bands on SDS PAGE gel

Dear all
sometimes i get convex shape of sample bands while running SDS PAGE.
Is it due to change in pH of buffers used in SDS PAGE gel prepn?

Thanks
ssacket
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Alt Today
Advertising
Google Adsense
 
This advertising will not be shown
in this way to registered members.
Register your free account today
and become a member on
Molecular Biology Forum
Standard Sponsored Links

  #2 (permalink)  
Old 05-28-2007, 04:14 PM
Pipette Filler
Points: 712, Level: 14Points: 712, Level: 14Points: 712, Level: 14
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Apr 2007
Posts: 5
mara1412 RSS Feed
Default Re: Convex shape of sample bands on SDS PAGE gel

Hi!
Sometimes convex shape results from gel overloading (usually I don't load more than 50 ug of protein for each lane), or you should run your gel slower than 100 V.
I never had problems with buffers.....but I store them always at +4°C.
Good luck!
m.
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
  #3 (permalink)  
Old 10-30-2007, 11:59 PM
Pipette Filler
Points: 325, Level: 6Points: 325, Level: 6Points: 325, Level: 6
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Oct 2007
Posts: 1
BG_Lung RSS Feed
Default Re: Convex shape of sample bands on SDS PAGE gel

I had the same issue (convex bands) in my western blot.

I did load 50ug-60ug protein, and only 2 of 8 lanes resulted in convex shape.
I ran it at 100V.

Overloading could have been the reason, but it is unclear since only 2 of the 8 bands had convex shape.

Any other suggestions anyone?

Thanks!!
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
  #4 (permalink)  
Old 10-31-2007, 06:33 PM
admin's Avatar
Administrator
Points: 8,891, Level: 65Points: 8,891, Level: 65Points: 8,891, Level: 65
Activity: 100%Activity: 100%Activity: 100%
 
Join Date: Nov 2005
Posts: 889
Blog Entries: 3
admin RSS Feed
Default Re: Convex shape of sample bands on SDS PAGE gel

Hello,
also you should make sure you load loading buffer in each lane including the ladder ( protein marker) as salt concentrations can also affect the sharpness and flatness of your bands.

Cheers!
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
  #5 (permalink)  
Old 12-04-2007, 12:09 AM
Pipette Filler
Points: 661, Level: 13Points: 661, Level: 13Points: 661, Level: 13
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Jun 2007
Posts: 7
Mossaded RSS Feed
Default Re: Convex shape of sample bands on SDS PAGE gel

That usually happens when you run your gels at too high of a voltage. It can also be due to your migration buffer so make sure your buffer is well mixed and homogenous. In my case, it usually evens out before it reaches the bottom so it doesn't cause any problems.
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Reply

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
Increase Exposure, Bands Disappear?? Mediatorx Western Blot Forum 3 07-01-2008 09:18 PM
SDS-PAGE Protocol moleculardude Protein Science 8 03-20-2008 03:23 PM
No Bands on Western Blot clokke76 Western Blot Forum 2 08-27-2007 04:26 PM


All times are GMT. The time now is 04:16 PM.


Powered by vBulletin® Version 3.7.1
Copyright ©2000 - 2008, Jelsoft Enterprises Ltd.
Copyright 2005-2007 Molecular Station | All Rights Reserved