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!
- - - - -

Protein measurements in 2% Triton X samples

  • Please log in to reply
2 replies to this topic

#1 Fribytter



  • Active Members
  • Pip
  • 6 posts

Posted 11 January 2012 - 08:22 AM

Hi all,

do any of you know what techniques can be used to determine protein quantity in cell lysates that contain 2% triton x. I have lysed my cells in lysis buffer containing 2% triton, because I am looking at a membrane bound protein (LC3) by WB.

Bradford etc. reagents are reacting very! good with Triton X, and the reagent gets saturated very fast by Triton.

Any suggestions?

#2 mdfenko


    an elder emeritus

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 3,266 posts

Posted 12 January 2012 - 08:22 AM

the bca protein assay is compatible with up to 5% triton x-100 (1% other tritons)

Edited by mdfenko, 12 January 2012 - 08:24 AM.

talent does what it can
genius does what it must
i do what i get paid to do

#3 JeremyDNA



  • Members
  • Pip
  • 2 posts

Posted 12 November 2013 - 07:37 PM

I have observed similar interference using a 2% Triton x-100 lysis buffer with Bradford assay.

Because triton reacts with coomassie blue (Bradford reagent), I ran a serial dilution of triton x-100 while keeping the rest of the buffer the same (no protein). The serial dilution was the following:

2%, 1%, 0.5%, 0.25%, 0.125%, 0.0625%, 0.03125%, 0.0156%, and 0% triton x-100 (in HBSS).

I found that loss of interference by triton x occurred somewhere near 0.0625%, which could explain why many lysis buffer recipes only use 0.05% triton x (presumably because their designers were using the Bradford assay)

Thus if you find sufficient yield with as low ad 0.05% triton, the Bradford is a suitable quantification technique.

Edited by JeremyDNA, 12 November 2013 - 09:09 PM.

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

©1999-2013 Protocol Online, All rights reserved.