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
- - - - -

Dot blot- great, western blot - no signal


  • Please log in to reply
3 replies to this topic

#1 Alsai

Alsai

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 21 April 2010 - 11:44 PM

Hi.

I'm working on MMPs. When I did dot blots I recived great signal but when I tried when I tried western blots there was no signal at all. I don't know whats wrong cause I'm using the same conditions as in dot blots. I checked my transfer and it's ok.

What else might be wrong?

#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,733 posts
125
Excellent

Posted 23 April 2010 - 12:09 PM

dot blots are usually of the native protein, western blots are of denatured protein. your antibody may not work with the denatured protein. if it is a commercial antibody, does the data sheet say that the antibody is certified for western blot?
talent does what it can
genius does what it must
i do what i get paid to do

#3 Alsai

Alsai

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 27 April 2010 - 04:17 AM

Yes, antibodies are for western blot.

dot blots are usually of the native protein, western blots are of denatured protein. your antibody may not work with the denatured protein. if it is a commercial antibody, does the data sheet say that the antibody is certified for western blot?



#4 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,733 posts
125
Excellent

Posted 27 April 2010 - 07:16 AM

the amount of protein in the band may be significantly lower than in the dot blot. you may need to incubate longer with and/or use a higher concentration of the antibody.
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.