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.
- 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
Mix reagent A and reagent B in a ratio of 50:1 (prepare at time of use).
Prepare a series of dilutions of a standard protein such as albumin.
Quantitation using test tubes
- Add 0.1 mL of the standard solution or a sample to the test tube.
- Add 2 mL of the prepared Working Solution to each tube and mix well.
- 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
- Bring the test tube to room temperature.
- Measure the absorbance at 562 nm using a spectrophotometer.
Quantitation using 96-well microplates
- Add 25 μL of either the standard solution or a sample to each well.
- Add 200 μL of the prepared Working Solution to each well and mix well.
- Allow them to react at 37 °C for 30 minutes.
- Bring the microplate back to room temperature.
- 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.
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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