Kit de ensayo de proteínas Solarbio BCA

La BCA (Ácido bicinconínico) Protein Assay Kit is a widely used method for the quantification of protein concentrations in a sample. This assay relies on the colorimetric detection of copper(II) ions reduced by proteins under alkaline conditions. The intensity of the resulting purple color is proportional to the protein concentration, allowing for precise quantification.

Here is a general guide on how to use a Proteína BCA Kit de ensayo:

Materiales necesarios:

1. Kit de ensayo de proteína BCA
2. Protein samples (standard solutions or unknown samples)
3. Microplaca o cubetas
4. Micropipetas y puntas
5. Lector de microplacas o espectrofotómetro
6. Agua destilada

Procedimiento:

1. Preparación de estándares de proteínas:
   • Prepare una serie de estándares de proteínas con concentraciones conocidas.. Estos estándares se utilizarán para crear una curva estándar para la cuantificación..
2. Preparation of Samples:
   • Dilute your protein samples to a concentration within the range of the standards. It’s important to have samples within the linear range of the assay for accurate quantification.
3. Preparación del reactivo de trabajo BCA:
   • Mix the BCA Reagent A (Bicinchoninic Acid solution) and BCA Reagent B (Copper(II) sulfate solution) in a 50:1 relación. Asegúrese de mezclar bien.
4. Incubación:
   • Add the BCA Working Reagent to your standards and samples, and then incubate the reaction mixture at a suitable temperature (commonly 37°C) para 30 minutos. Durante este tiempo, proteins in the samples will reduce the copper ions, forming a purple-colored complex.
5. Medición:
   • After incubation, measure the absorbance of each well at a wavelength around 562 nm usando un lector de microplacas o espectrofotómetro. The absorbance is directly proportional to the protein concentration.
6. Creación de curva estándar:
   • Trazar los valores de absorbancia de los estándares frente a sus concentraciones conocidas para crear una curva estándar.. This curve will be used to determine the protein concentrations of unknown samples.
7. Quantification of Unknown Samples:
   • Measure the absorbance of your unknown samples and use the standard curve to determine their protein concentrations.
8. Análisis de datos:
   • Calculate the protein concentrations based on the standard curve. Most microplate readers have software that can automate this process.
9. Resultados:
   • Report the protein concentrations of your samples in the appropriate units (p.ej., mg/mL) based on the BCA assay.

It’s important to follow the specific instructions provided by the manufacturer of the BCA Protein Assay Kit, as the exact protocol may vary between kits. Además, ensure that all reagents are properly stored, and the assay is performed under the recommended conditions for accurate and reliable results.

The BCA Protein Assay Kit is based on the protein reduces Cu2+ to Cu+ and the principle that BCA forms a purple-blue complex with Cu1+ under alkaline conditions, followed by detecting absorbance at 562nm and comparing with the standard curve to quantify the protein.