Free Beer-Lambert Protein Calculator

Calculate protein concentration from UV absorbance at 280 nm using the Beer-Lambert law and known extinction coefficient.

M⁻¹cm⁻¹
cm
Da

Mass Concentration

0.0010 mg/mL

Molar Concentration0.00002326 M

Mass Concentration vs Absorbance at 280 nm

Protein Concentration from A280

UV absorbance at 280 nm is widely used to measure protein concentration. Aromatic residues (Trp, Tyr) and disulfide bonds absorb at this wavelength.

Formula

c (M) = A280 / (epsilon × l)

c (mg/mL) = c (M) × MW / 1000

The extinction coefficient epsilon can be calculated from the amino acid sequence using tools like ProtParam. Typical values range from 10,000 to 100,000 M⁻¹cm⁻¹.

Example Calculation

A protein with epsilon = 43,000 M⁻¹cm⁻¹ and MW = 45 kDa, measured A280 = 1.0 in a 1 cm cuvette.

  1. 01c = 1.0 / (43000 × 1) = 2.326 × 10⁻⁵ M = 23.26 uM
  2. 02c = 23.26 × 10⁻⁶ × 45000 / 1000 = 1.047 mg/mL

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