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!

yobou's Content

There have been 2 items by yobou (Search limited from 26-January 19)


Sort by                Order  

#182096 how to enhance the detection limit of home-made Bradford reagent?

Posted by yobou on 17 January 2020 - 03:45 AM in Protein and Proteomics

Dear All

I have prepared Bradford reagent as follows:

 

  1. Dissolve 50mg of Coomassie Blue G250 in 50ml of methanol.
  2. Add 100ml of 85% H3PO4 to the solution from step 1.
  3. Add the solution from step 2 into 500ml of H2O and mix.
  4. Filter to remove and precipitates.
  5. Add an additional 350ml of H2O.
  6. Store at 4oC.

 

HOWEVER, this prepared reagent cannot distinguish between low protein concentrations (0.5 and 0.1 mg/mL) where both concentrations give the same absorbance values. 

HOW can I enhance the detection limit of the prepared BRADFORD to be comparable to the commercial one purchased from Bio-Rad?

 

Since I am using lysis buffer that contains detegents like SDS or NP40, igpal. I need to know if there is a special modification to Bradford reagent.

Thanks a lot




#182095 how to enhance the detection limit of home-made Bradford reagent?

Posted by yobou on 16 January 2020 - 02:39 AM in Protein and Proteomics

Dear All

I have prepared Bradford reagent as follows:

 

  1. Dissolve 50mg of Coomassie Blue G250 in 50ml of methanol.
  2. Add 100ml of 85% H3PO4 to the solution from step 1.
  3. Add the solution from step 2 into 500ml of H2O and mix.
  4. Filter to remove and precipitates.
  5. Add an additional 350ml of H2O.
  6. Store at 4oC.

 

HOWEVER, this prepared reagent cannot distinguish between low protein concentrations (0.5 and 0.1 mg/mL) where both concentrations give the same absorbance values. 

HOW can I enhance the detection limit of the prepared BRADFORD to be comparable to the commercial one purchased from Bio-Rad?





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

©1999-2013 Protocol Online, All rights reserved.