Protein Concentration (Bradford) Calculator Formula

Understand the math behind the protein concentration (bradford) calculator. Each variable explained with a worked example.

Formulas Used

Protein Concentration

conc_undiluted = (absorbance - intercept) / slope * dilution

Protein Concentration

conc_mg = (absorbance - intercept) / slope * dilution / 1000

Variables

VariableDescriptionDefault
absorbanceAbsorbance at 595 nm0.45
slopeCalibration Slope (m)0.05
interceptCalibration Intercept (b)0.02
dilutionDilution Factor1

How It Works

Bradford Protein Assay

The Bradford assay uses Coomassie Brilliant Blue G-250 dye, which shifts from red to blue upon binding protein. Absorbance at 595 nm is proportional to protein concentration.

Calculation

[Protein] = (A595 - b) / m × dilution factor

where m and b come from a standard curve using BSA or another reference protein. The Bradford assay is sensitive to 1-20 ug/mL but has protein-to-protein variation.

Worked Example

A Bradford assay reads A595 = 0.45, standard curve: m = 0.05, b = 0.02, no dilution.

absorbance = 0.45slope = 0.05intercept = 0.02dilution = 1
  1. 01[Protein] = (0.45 - 0.02) / 0.05 × 1
  2. 02[Protein] = 0.43 / 0.05 = 8.60 ug/mL