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
- - - - -

HA-TAG


  • Please log in to reply
2 replies to this topic

#1 mikew

mikew

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 80 posts
0
Neutral

Posted 21 October 2009 - 10:49 AM

Hi,
I have HA tagged my protein. My sequencing reaction looks good. Everything should be in frame.
I have added a single HA tag.
The problem is that my protein is expressed (by Western) but I cannot detect it with an HA tag antibody (I use a 3x HA tag for control).
Any ideas?

#2 fred197237

fred197237

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 13 November 2009 - 12:58 PM

the HA tag protein is so small ~ 1-2 kd so might need to use a suitable gel and markers

#3 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,706 posts
567
Excellent

Posted 15 November 2009 - 04:01 PM

Hmmm, I had a similar problem not long ago... it was the transfection that wasn't working.

My single HA tag works just fine, I am using a Sigma anti-HA rabbit antibody, which shows a strong band of the right size and a much weaker and slightly smaller band in the controls and treatments.

Is there a spacer between the HA and the protein of interest? If not, direct linkage has been known to affect the binding of antibodies to tags.

Edited by bob1, 15 November 2009 - 04:01 PM.





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

©1999-2013 Protocol Online, All rights reserved.