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
- - - - -

Tagged recombinant protein has different mobility?

tagged protein

  • Please log in to reply
4 replies to this topic

#1 An-nur Al-Irsyad

An-nur Al-Irsyad

    member

  • Active Members
  • Pip
  • 5 posts
2
Neutral

Posted 28 November 2012 - 01:30 AM

I did an expression of Adh gene in BL21 (DE3). I purified the protein using histidine tag column. Then, I run the total protein and also purified Adh protein in same SDS PAGE gel. BUT, the purified histidine tagged protein has a quite larger size than expected size. anyone can explain why this happen?Is it tagged recombinant protein has different mobility? anyone can explain this to me?

Edited by An-nur Al-Irsyad, 28 November 2012 - 01:31 AM.


#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,236 posts
336
Excellent

Posted 28 November 2012 - 12:08 PM

It is relatively common for proteins not to migrate at their expected sizes. Adding a tag could easily alter the charge on the protein, and thereby alter how it runs. SDS PAGE gels are not totally based on protein size, with the SDS creating the all of the charge; PI and a few other factors also play a role.

#3 An-nur Al-Irsyad

An-nur Al-Irsyad

    member

  • Active Members
  • Pip
  • 5 posts
2
Neutral

Posted 22 January 2013 - 10:12 PM

SDS is detergent that give negative charge on protein-correct me if im wrong..
I though only native page rely on the protein charge, pI Posted Image

#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,236 posts
336
Excellent

Posted 23 January 2013 - 12:15 AM

SDS is detergent that give negative charge on protein-correct me if im wrong..
I though only native page rely on the protein charge, pI Posted Image

What you say is true. Native PAGE does only rely on charge and size, but SDS-PAGE also has some component of charge migration, despite the SDS.

#5 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,612 posts
118
Excellent

Posted 23 January 2013 - 06:59 AM

this may give you some insight:

Attached Files


talent does what it can
genius does what it must
i do what i get paid to do




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

©1999-2013 Protocol Online, All rights reserved.