Solarbio BCA Protein Assay Kit 50T

The BCA (Bicinchoninic Acid) Protein Assay Kit is a widely utilized colorimetric method for the quantification of protein concentrations in a sample. This assay relies on the reduction of Cu^2+ ions to Cu^1+ by proteins under alkaline conditions, forming a purple-colored complex with bicinchoninic acid. The intensity of the color is directly proportional to the protein concentration, enabling accurate quantification.

Here is a step-by-step guide on how to use a BCA Protein Assay Kit:

Materials Required:

1. BCA Protein Assay Kit
2. Protein standards (bovine serum albumin is commonly used)
3. Sample containing unknown protein concentrations
4. Microplate or cuvettes
5. Micropipettes and tips
6. Microplate reader or spectrophotometer
7. Distilled water

Procedure:

1. Preparation of Protein Standards:
   • Prepare a series of protein standards with known concentrations. These standards will be used to create a standard curve for quantification. Common concentrations are 0, 0.125, 0.25, 0.5, 0.75, and 1.0 mg/mL.
2. Preparation of Sample:
   • Dilute your protein sample to a suitable concentration within the linear range of the assay. If needed, you may need to perform a preliminary dilution to ensure that the sample falls within the standard curve.
3. Preparation of BCA Working Reagent:
   • Mix BCA Reagent A and BCA Reagent B in a 50:1 ratio (typically provided in the kit). Ensure thorough mixing.
4. Addition of Standards and Sample:
   • Pipette aliquots of the protein standards and the sample into separate wells of a microplate. Each standard and sample should be run in duplicate or triplicate.
5. Addition of BCA Working Reagent:
   • Add an appropriate volume of the BCA Working Reagent to each well containing standards and samples. Mix thoroughly.
6. Incubation:
   • Incubate the microplate at an appropriate temperature (typically 37°C) for 30 minutes to allow the color reaction to occur.
7. Measurement:
   • Measure the absorbance of each well at 562 nm using a microplate reader or spectrophotometer.
8. Creation of Standard Curve:
   • Plot the absorbance values of the standards against their known concentrations to create a standard curve.
9. Quantification of Sample:
   • Use the standard curve to determine the protein concentration of your sample based on its absorbance.
10. Data Analysis:
    • Calculate the concentration of protein in the sample based on the standard curve. Most microplate readers have software that can assist in this analysis.

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