Protein Assay BCA Reagent

• 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.

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.

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.