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Protein Assay BCA Reagent

Protein Assay BCA Kit is a reagent kit for quantitative determination of protein levels in solution, using bicinchoninic acid.

Proteins reduce cupric cation (Cu2+) to cuprous cation (Cu+) under alkaline conditions. The Cu+ forms a chelate complex with bicinchoninic acid. The solution containing the chelate complex turns a reddish purple color. Protein concentrations can be determined by measuring the absorbance at 562 nm.

Kit Components

  • Reagent A: BCA solution 500 mL x 1 bottle
  • Reagent B: Copper(Ⅱ) sulfate solution 25 mL x 1 bottle

* This kit does not include albumin standard solution.

Measurement Method

Preparation of reagent

Working solution

Mix reagent A and reagent B in a ratio of 50:1 (prepare at time of use).

Standard solution

Prepare a series of dilutions of a standard protein such as albumin.

Quantitation using test tubes

  1. Add 0.1 mL of the standard solution or a sample to the test tube.
  2. Add 2 mL of the prepared Working Solution to each tube and mix well.
  3. Allow them to react under the conditions described below.
    Reaction conditions Quantitative range
    Standard method 37 ℃, 30 minutes 20~2,000 μg/mL
    Room temperature method Room temperature, 2 hours 20~2,000 μg/mL
    High-sensitivity method 60 ℃, 30 minutes 5~250 μg/mL
  4. Bring the test tube to room temperature.
  5. Measure the absorbance at 562 nm using a spectrophotometer.

Quantitation using 96-well microplates

  1. Add 25 μL of either the standard solution or a sample to each well.
  2. Add 200 μL of the prepared Working Solution to each well and mix well.
  3. Allow them to react at 37 °C for 30 minutes.
  4. Bring the microplate back to room temperature.
  5. Measure the absorbance at 562 nm using a microplate reader.

Measurement Example

After mixing reagents A and B in the ratio of 50:1, mix the combination with 31.25, 62.5, 125, 250 μg/mL of BSA solution in the ratio of 20:1, in test tubes. The absorbance of the test tubes is measured with a high-sensitivity method.

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Protein Determination Reagents

Protein Standard

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