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
- - - - -

Primary antibody


  • Please log in to reply
1 reply to this topic

#1 manojdarolia

manojdarolia

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 22 November 2004 - 11:20 PM

well i want to know the minimum time requried to incubate the membrane with primary antibody while doing western and the stringency of the the washes i mean washing time and washing buffer volume,

can anybody tell me about that?

thanks

#2 Sprag

Sprag

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 45 posts
0
Neutral

Posted 25 November 2004 - 09:37 AM

well, the minimum time for primary can be as low as 20min, but i do recommend 60min. The washes, I do 3 times 20min in a 30-50ml for a minigel, but you can do shorter times, depends on your antibody. Nice western is a balance between high signal and low background, so if you get no background at all, you can easily shorten your incubation times, whereas higher background, you can either do longer washes, higher salt/tween/milk, or you can lower your concentration of primary...

have fun,




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

©1999-2013 Protocol Online, All rights reserved.