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
- - - - -

Bradford assay+high urea


  • Please log in to reply
3 replies to this topic

#1 bluebird

bluebird

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 33 posts
0
Neutral

Posted 28 October 2009 - 04:46 AM

I have protein solublised in 8M urea buffer, do you I can apply bradford assay for protein solublised in high conc. of urea?
if it is OK do I need to prepare the standards from the same buffer that my protein is soulube in( 8Murea)? or I can use H2O?

Edited by bluebird, 28 October 2009 - 04:50 AM.


#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,706 posts
123
Excellent

Posted 28 October 2009 - 06:13 AM

bio-rad says that their reagent is compatible with 6M urea. that's undiluted sample in the standard assay. if you dilute or use a small volume (relative to the reagent volume) then you should be okay with 8M.

it is always better, but not always necessary, to run your standards in the same buffer as your sample (we use a buffer blank because we may be running samples with more than one buffer).

Edited by mdfenko, 28 October 2009 - 06:14 AM.

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

#3 bluebird

bluebird

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 33 posts
0
Neutral

Posted 28 October 2009 - 06:34 AM

I will use a 96 well and I will pipette 0,2,4...15 and 20 micL of BSA STANDARD solution and make up with ddH2O 20micL , and the sample is 20micL , and 200micL bradford reagent. Do you think the ratio between the reagent and the buffer is relatively small in this protocol?

#4 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,706 posts
123
Excellent

Posted 28 October 2009 - 10:16 AM

the standard assay is 4:1 reagent:sample, yours is 10:1. it should be okay. try it.
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.