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.


Centrifuge after sonication

Centrifuge after sonication - Western Blot Forum

Centrifuge after sonication - 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 12-31-2009, 02:20 PM
Pipette Filler
Points: 501, Level: 10 Points: 501, Level: 10 Points: 501, Level: 10
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Aug 2009
Posts: 6
Thanks: 0
Thanked 0 Times in 0 Posts
Default Centrifuge after sonication



Hi everyone,

I collect whole hippocampi from mice then sonicate the samples in a lysis buffer which contains 50 mM Tris (pH 7.4), 1 mM EDTA, 1 mM EGTA, 1% SDS, and a protein/phosphatase inhibitor cocktail. I've seen some protocols which then suggest to centrifuge the samples after sonication. I was wondering what was the purpose of centrifuging the samples after sonication? I was hoping that the community here at Molecular Station would be able to provide me as much information/answers as possible. Thanks!
Reply With Quote
  #2  
Old 10-09-2012, 01:39 PM
Pipette Filler
Points: 324, Level: 6 Points: 324, Level: 6 Points: 324, Level: 6
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2012
Posts: 10
Thanks: 6
Thanked 2 Times in 2 Posts
Default Re: Centrifuge after sonication

As far as I know, Lysis buffer is going to break cell wall, so DNA will be free and then, it will be sonicated. The spin after sonication is usually a high-speed centrifugation during few minutes, which allows to precipitate all debris of your samples. It's like the last step to discard organelles, cell wall, etc.
If you want, you can add another step. After centrigufation, you can filter the sonicated sample through 45um syringe or miracloth membrane to get a clean lysate.

I hope it would be useful!
Reply With Quote
Reply

Tags
centrifuge , sonication


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
Selling a Centrifuge sucktide Lab Equipment Discussion and Reviews 1 01-28-2010 10:00 AM
High performance Floor Centrifuge suggestions badgtx Lab Equipment Discussion and Reviews 1 12-03-2009 06:35 PM
Hettich centrifuge schematics James Herbert Protein Forum 0 09-21-2009 12:42 PM
problem in centrifuge of protein waffaa_qm Protein Science 0 09-20-2007 12:03 AM
[Cell-biology] old centrifuge without a speed indicator Jayakumar, R Cell Biology and Cell Culture 0 04-19-2007 06:17 PM


All times are GMT. The time now is 11:37 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 0.11704 seconds with 16 queries