Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

protein purification


  • Please log in to reply
3 replies to this topic

#1 eman

eman

    member

  • Members
  • Pip
  • 4 posts
-1
Neutral

Posted 21 December 2010 - 12:02 PM

i want to ask about how can i make purification to my protein? the M.wt of the protein is 5kDa in expression vector PET-29 in Bl21 bacteria

#2 protolder

protolder

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 293 posts
9
Neutral

Posted 21 December 2010 - 11:05 PM

Hola, in pET 29 you can have your protein fused with Nt6His,-is your case?.If yes, after lysis of bacterial pellet I would use any of the commercial resins of Ni++ or Co++ with th addecuate device for the volume of culture, spin columns, FPLC colums, or resin in batch under the conditions of the supplier. Buena suerte

#3 BioMiha

BioMiha

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 229 posts
9
Neutral

Posted 21 December 2010 - 11:06 PM

If you are expressing your protein in E. coli the best way to purify it is by attaching a His tag (or any other tag). Otherwise you can use a number of methods depending on the characteristics of your protein. There is no pre-defined way to purify a protein, it all depends on what the protein is like, what the sample is like, how clean you need it to be, what you need it for, etc., etc...

#4 Allenwood

Allenwood

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 03 January 2011 - 01:32 AM

Nice information........

thanks.




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.