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
- - - - -

ELISA Extraction Buffer for Plants?


  • Please log in to reply
4 replies to this topic

#1 WolfeMD

WolfeMD

    member

  • Active Members
  • Pip
  • 18 posts
0
Neutral

Posted 06 April 2009 - 01:32 PM

Can anyone recommend a good extraction buffer that is compatible with ELISA that will work for Arabidopsis thaliana?

I have previously used SDS based buffers to do Western Blots, but SDS isn't compatible with ELISA because it denatures antibodies. I have had extremely limited success trying to use PBST (phosphate buffered saline with Tween-20).

Any advice would be helpful.

Cheers,

Marnin

#2 sgt4boston

sgt4boston

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 276 posts
1
Neutral

Posted 09 April 2009 - 11:45 AM

After you do the extraction could you not change buffers by dialysis?

#3 WolfeMD

WolfeMD

    member

  • Active Members
  • Pip
  • 18 posts
0
Neutral

Posted 17 April 2009 - 07:20 AM

After you do the extraction could you not change buffers by dialysis?


I could potentially do that. However, that is an entire extra step. I would only do that if it is the only way. Additionally, extra steps like that make ELISA less quantitative because there are extra steps with extra opportunity for human error.

Does ANYONE work on plants here?

Help!?

#4 sgt4boston

sgt4boston

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 276 posts
1
Neutral

Posted 22 April 2009 - 08:30 AM

Could you extract; concentrate your material of interest using a Milllipore product (low binding) then dilute your sample? Depending upon the degree of concentration the % of surfactant should be fairly low and not interfere in the test.

#5 scary

scary

    member

  • Active Members
  • Pip
  • 11 posts
0
Neutral

Posted 28 May 2009 - 12:49 AM

Hi,

I don't work with arabidopsis but I work on some crop plants like Groundnut, Cotton, Pigeon pea, Sunflower where I use PBST or TBST @ pH-7.6 or 50mM Tris at pH-8.0 as ELISA extraction buffers. Check with these buffers it may help you.

All the best

Scary




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

©1999-2013 Protocol Online, All rights reserved.