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
- - - - -

problems with negative control


  • Please log in to reply
2 replies to this topic

#1 michalcn

michalcn

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 32 posts
0
Neutral

Posted 11 July 2011 - 07:01 PM

Hi,
I did co-ip with mouse ab and mouse IgG as negative control.blot it and prob with different ab but again mouse.
I'm getting very nice band at the expected size compare to the control, but i can also see it, less strong, in the igG
is this still can be positive and how can I reduce it
Thanks
Michal

#2 GNANA

GNANA

    GNANA

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 209 posts
9
Neutral

Posted 12 July 2011 - 01:08 AM

Have you done the Preclearing with the Beads (Protein G i guess for Mouse) ?? this would reduce the background in Negtive control..Try out this and see if you havent done. and increase washing steps, this might also help...

.

Hi,
I did co-ip with mouse ab and mouse IgG as negative control.blot it and prob with different ab but again mouse.
I'm getting very nice band at the expected size compare to the control, but i can also see it, less strong, in the igG
is this still can be positive and how can I reduce it
Thanks
Michal


I would prefer being perfectionist rather than a passionist in Research.

I always had an alternate hypothesis....

#3 michalcn

michalcn

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 32 posts
0
Neutral

Posted 12 July 2011 - 06:00 PM

I did the precleaning with the beads and increased the washes without change,
I'm using the protein a/g agarose beads from santa cruz
I heard that there is secondary ab that do not recognize the Igg, does anyone know what does it mean?
Thanks
M

Have you done the Preclearing with the Beads (Protein G i guess for Mouse) ?? this would reduce the background in Negtive control..Try out this and see if you havent done. and increase washing steps, this might also help...

.


Hi,
I did co-ip with mouse ab and mouse IgG as negative control.blot it and prob with different ab but again mouse.
I'm getting very nice band at the expected size compare to the control, but i can also see it, less strong, in the igG
is this still can be positive and how can I reduce it
Thanks
Michal






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

©1999-2013 Protocol Online, All rights reserved.