The standard Pierce BCA Protein Assay detects protein concentrations from 20 to 2,000 μg/mL using a two-component system: Reagent A, a carbonate buffer containing BCA reagent, and Reagent B, a cupric sulfate solution, which are combined to make an apple green–colored working solution that turns purple after 30 minutes at 37°C in the presence of protein. Shortly before the end of the incubation period, the cells are evaluated microscopically. Assessment of growth inhibition of cells by extraction method and BCA Staining The test item is extracted under agitation for a defined period of time in DMEM supplemented with 10% FBS at 37 ± 1°C and L929 cells are incubated with the extract. This procedure is very applicable to microtiter plate methods. The BCA is used for the same reasons the Lowry is used. The bicinchoninic acid (BCA) assay is available in kit form from Pierce (Rockford, Ill.). The product of this reaction is a cupric ion that is coordinated to peptide nitrogen atoms. Stoscheck (1990) has suggested that the BCA assay will replace the Lowry because it requires a single step, and the color reagent is stable under alkaline conditions. The standard Pierce BCA Protein Assay detects protein concentrations from 20 to 2,000 μg/mL using a two-component system: Reagent A, a carbonate buffer containing BCA reagent, and Reagent B, a cupric sulfate solution, which are combined to make an apple green–colored working solution that turns purple after 30 minutes at 37°C in the presence of protein. BCA-Assay, Bicinchoninsäure-Assay, eine Methode zur quantitativen Proteinbestimmung, die eine Kombination der Biuret-Reaktion mit Bicinchoninsäure (BCA) als Detektionssystem darstellt. It is based off of the Biuret reaction which involves the coordination between the peptide backbone and cuprous ion under alkaline conditions (pH 11.25). The Bicinchoninic Assay (BCA) is a contemporary method for determining protein concentration.


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