Bradford Assay Biology Virtual Lab Simulation| PraxiLabs

Bradford Assay

Biology | Biochemistry | Proteomics

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Bradford Assay Virtual Lab Simulation

Importance of Bradford Assay Virtual Lab Simulation

To measure the concentration of proteins in a solution by colorimetric analysis using Bradford assay.

Bradford Assay Virtual Lab Simulation Method

Protein concentration measurement according to Bradford assay method.

Objectives of Bradford Assay Virtual Lab Simulation

  • <p>Identify the amino acids that the Bradford Protein Assay measures.

  • Describe the color change that occurs when proteins combine with Coomassie dye under acidic conditions.

  • Perform serial dilutions of standard.

  • Illustrate the correct standard curve equation for an example BSA standard.

  • Interpret the standard curve equation when given example data.

  • Calculate the protein concentration of an example. Recall the substance commonly used as standards in the assay and the device used to measure the color of the samples.</p>

Theory of Bradford Assay Virtual Lab Simulation

Colorimetric assays are methods to measure protein concentration in samples. They are based on color changes that occur when the protein in the sample interacts with a specific reagent causing color changes that can be measured using a spectrophotometer. Commonly used assays are the Bradford assay, the Lowry assay and the BCA assay.
The Bradford assay is considered a protein concentration measurement experiment based on the binding of the basic amino acids of a protein to a dye present in Bradford reagent (coomassie brilliant blue). This leads to a shift in the absorbance maximum of the dye from 465 to 595
Bradford reagent contains the coomasie blue dye. The cationic (unbound) form is green / red with an absorption maximum at 465 nm. The anionic bound form of the dye, held together by hydrophobic interactions, has an absorption maximum at 595 nm. Thus; the increase of absorbance from 465 to 595 nm is directly proportional to the amount (concentration) of protein bound to the dye.

Bradford Assay Principle of Work

Bradford assay virtual lab simulation is based on the binding of the basic amino acids of a protein to a dye present in Bradford reagent (e.g. Coomassie brilliant blue). This leads to a shift in the absorbance maximum of the dye from 465 to 595.*After creating a standard curve of protein solutions with known concentrations, the protein concentration of unknown samples can be plotted on the standard dilution curve and calculated.

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