Solution preparation

Bradford Stock Solution Preparation

This solution is mainly used to determine the concentration of proteins in solution.

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MN Editors
This article writter by MN Editors on May 04, 2021

Microbiology Notes is an educational niche blog related to microbiology (bacteriology, virology, parasitology, mycology, immunology, molecular biology, biochemistry, etc.) and different branches of biology.

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Bradford Stock Solution Preparation
Bradford Stock Solution Preparation

Bradford Stock Solution

  • This solution is mainly used to determine the concentration of proteins in solution.
  • The principle of detection is based on the formation of a complex between the dye, Brilliant Blue G, and proteins in solution.
  • The protein dye complex causes a shift in the absorption maximum of the dye from 465 to 595 nm. The amount of absorption is proportional to the protein present.
  • The Bradford Reagent requires no dilution and is suitable for micro, multiwell plate, and standard assays. 
  • This solution also compatible with reducing agents which are often used to stabilize proteins in solution.
  • This solution is consist of t consists of Brilliant Blue G in phosphoric acid and methanol.
  • Bradford Stock Solution is stable at 2–8 °C in an unopened container for at least 1 year.

Preparation of Bradford Stock Solution

To prepare 1L of Bradford Stock Solution, the following components are required;

ComponentAmountConcentration
Ethanol (95%) (mw: 46.07 g/mol)333 mL31.67 %
Phosphoric acid (88%) (mw: 58.4 g/mol)667 mL58.67 %
Coomassie Brilliant Blue (mw: 825.97 g/mol)1.16 g1.41 M

Procedure

  1. Prepare 0 mL of distilled water in a suitable container.
  2. Add 333 mL of Ethanol (95%) to the solution.
  3. Add 667 mL of Phosphoric acid (88%) to the solution.
  4. Add 1.16 g of Coomassie Brilliant Blue to the solution.
  5. Add distilled water until volume is 1 L.

References

  • “Bradford Stock Solution Preparation.” AAT Bioquest, Inc, 04 May. 2021, https://www.aatbio.com/resources/buffer-preparations-and-recipes/bradford-stock-solution.
  • https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Bulletin/b6916bul.pdf

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Microbiology Notes is an educational niche blog related to microbiology (bacteriology, virology, parasitology, mycology, immunology, molecular biology, biochemistry, etc.) and different branches of biology.

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