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!
- - - - -

Bradford Protein Assay - Problem in BSA Standard Curve

Bradford Protein Assay

  • Please log in to reply
2 replies to this topic

#1 bamya



  • Members
  • Pip
  • 1 posts

Posted 07 August 2012 - 05:18 AM

I am trying to determine protein concentration but my standard curve does not look like a standard curve at all.
I used to use BSA stock prepared by distilled water but I got high protein concentration all the time.
Then my advisor suggested me to use RIPA instead of water. So I prepared a BSA stock prepared by 0.5mg BSA powder and 10 ml RIPA lysis buffer (10mg BSA powder/200 ml RIPA). I put 25ul, 50 ul, 75 ul of my stock and so on into each well. Then I used PBS for both my standard curve samples and unknown samples to complete 200ul (i am using 96-well plate). But this time detergent and Bradford Assay reagent precipitated.
I have to find a solution on my own. If anyone can suggest me a solution I will be very happy. I hope I can explain the steps I have done until now clearly.
Thanks in advance

#2 mdfenko


    an elder emeritus

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 3,268 posts

Posted 07 August 2012 - 08:30 AM

we make our stocks and dilutions with water. then we prepare subtractive blanks with the buffer(s) in which our samples reside.
talent does what it can
genius does what it must
i do what i get paid to do

#3 bob1


    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,511 posts

Posted 07 August 2012 - 02:07 PM

Check your bradford solutions haven't gone off, they are relatively unstable so may well have precipitated.

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

©1999-2013 Protocol Online, All rights reserved.