Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Proteomics Forum > 2-D Gel Electrophoresis Forum
Register Search Today's Posts Mark Forums Read

2-D Gel Electrophoresis Forum 2-D Gel Electrophoresis Forum. A forum dedicated to two dimensional gel electrophoresis, discussion about systems, and troubleshooting its problems.


Precipitating plant proteins

Precipitating plant proteins - 2-D Gel Electrophoresis Forum

Precipitating plant proteins - 2-D Gel Electrophoresis Forum. A forum dedicated to two dimensional gel electrophoresis, discussion about systems, and troubleshooting its problems.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 07-07-2009, 11:58 AM
Pipette Filler
Points: 456, Level: 9 Points: 456, Level: 9 Points: 456, Level: 9
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jun 2009
Posts: 4
Thanks: 0
Thanked 0 Times in 0 Posts
Angry Precipitating plant proteins



I am working with rhizome proteins i extract using a phenol based method. The proteins have given me good SDS PAGE bands. My problem now is that my proteins of late seem to be insoluble in urea lysis buffer (7M urea, 2M thiourea, 2% chaps) after hours of vortexing. Sometimes the proteins dissolve and i keep the solutin at -20C. Upon thawing, the proteins tend to precipita. I was planning to go to IEF and 2D electrophoresis, but with the problems i have highlighted, i am very worried. Please help me.

warm regards

Proteome
Reply With Quote
  #2  
Old 05-01-2013, 05:52 PM
Pipette Filler
Points: 56, Level: 1 Points: 56, Level: 1 Points: 56, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2013
Posts: 4
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Precipitating plant proteins

Don't let the sample stand for hrs.You should load the sample within 1hr after dissolving in buffer.
Try 10% TCA in Acetone precipitation method.
And after precipitation add rehydration buffer and let it be so for 10 min then dissolve sample slowly with an spatula.
Reply With Quote
Reply

Tags
plant , precipitating , proteins


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
Human Cytome Project - Update 24 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 1 08-01-2010 02:18 PM
Postdoctoral Research Associate position in Plant Molecular and CellBiology Reumann, Sigrun Arabidopsis and Plant Biology 0 12-21-2007 01:24 AM
Plant Science positions at the University of Nebraska Julie M. Stone Arabidopsis and Plant Biology 0 11-17-2007 01:36 PM
4 Post doc positions within Plant Biology at the University ofCopenhagen Søren Bak Arabidopsis and Plant Biology 0 03-25-2007 08:52 PM
Human Cytome Project - Update 6 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 0 01-06-2005 11:18 AM


All times are GMT. The time now is 06:50 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 0.15373 seconds with 16 queries