Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register 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.


Methods Digest, Vol 48, Issue 10

Methods Digest, Vol 48, Issue 10 - Protocols and Methods Forum

Methods Digest, Vol 48, Issue 10 - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 05-14-2009, 02:44 AM
TieQiao Wu
Guest
 
Posts: n/a
Default Methods Digest, Vol 48, Issue 10



Hi

Does anyone have some experiences to purify insoluble GST-fusion
protein? I tried to purify GST-fusion protein recently, but it looks
that the protein did not bind to the beads, I did coommassie stain and
western blotting and results show the protein was expressed. I purified
the same protein last year although the fusion protein degraded
partially. In the cell suspension 1M DTT and 10% Sarkosyl were added
before sonication, 10% Triton X-100 was added after sonication, the
final concentrations are: 0.7% Sarkosyl, 2% Triton X-100 and 5mM DTT.

Thanks a lot!

Tieqiao


This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed.
If you have received this email in error please promptly delete this email and notify the system manager.

This email has been scanned by the MessageLabs Email Security System.
For more information please visit [Only registered users see links. ]

Reply With Quote
  #2  
Old 05-15-2009, 04:28 PM
Dr Engelbert Buxbaum
Guest
 
Posts: n/a
Default Methods Digest, Vol 48, Issue 10

Am 13.05.2009, 22:44 Uhr, schrieb TieQiao Wu <[Only registered users see links. ].au>:


That can happen, and you have two options:
- use a differnet tag (or a different position of tag, N- vs C-terminal)
and hope that it will work
- do a conventional purification (IEC, SEC, HIC etc)
Reply With Quote
Reply

Tags
digest , issue , methods , vol


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
Methods Digest, Vol 49, Issue 12 TieQiao Wu Protocols and Methods Forum 0 06-14-2009 11:28 PM
Methods Digest, Vol 49, Issue 12 TieQiao Wu Protocols and Methods Forum 0 06-14-2009 11:28 PM
Methods Digest, Vol 34, Issue 23 Protocols and Methods Forum 1 03-27-2008 10:57 AM
Methods Digest, Vol 17, Issue 23 Sayyari, Mohammad Protocols and Methods Forum 0 10-25-2006 09:11 AM
Methods Digest, Vol 3, Issue 4 TieQiao Wu Protocols and Methods Forum 0 08-05-2005 01:03 AM


All times are GMT. The time now is 03:32 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 0.11934 seconds with 16 queries