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Protein Assays

Bradford Assay

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Learn how a Bradford Assay works and how it can be used to estimate the concentration of protein in a solution. Explore the properties of the dye Coomassie Brilliant Blue and how changes in the protein concentration affect the dye's absorbance. Practise preparing solutions for the assay and understand the errors introduced when sample absorbance falls outside the analytical range.

Learning Outcomes

  • Learn about the principles of the assay
  • Familiarise yourself with the different solutions measured in the assay
  • Explore the steps of the protocol and analysis and how best to obtain accurate results
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Concepts Principles and Theories

  • How a change in the charge, absorbance max and colour of the CBB dye upon protein binding forms the principle of the assay
  • How protein concentration affects the colour of the solution and the absorbance reading

Designing and Planning Experiments

  • How the components of the blank, standards and samples differ and why it is important to prepare all three

Obtaining and Recording Data

  • Why it is important to mix the solutions prior to measuring the absorbance
  • Why it is important to leave the solutions for 5 minutes prior to measuring the absorbance 
  • Why it is important to blank the spectrophotometer prior to measuring the absorbance 

Interpreting and Analysing Data

  • How the accuracy of the interpolation is affected by where the sample concentration falls within the assay range

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Protein Assays

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