A Bicinchoninic Acid Protein Assay Kit (BCA kit) (Sigma Aldrich, USA) was used for the determination of protein concentration in the prepared samples. For this purpose, a 1 : 50 mixture of copper sulphate solution (II) and bicinchoninic acid solution was prepared. 25 μl of the examined protein was added to 200 μl of the mixture, and incubated for 30 minutes at 37˚C. Next, the absorbency of the research sample against control sample was measured at wavelength λ = 550 nm. The protein concentration was determined based on a standard curve of bovine serum albumin in a concentration of 0.1-1 mg/ml.
Bicinchoninic acid protein assay kit bca kit
Manufactured by Merck Group
Sourced in United States
The Bicinchoninic Acid Protein Assay Kit (BCA kit) is a colorimetric detection and quantitation assay for determining the total protein concentration in a sample. The kit utilizes the bicinchoninic acid (BCA) reaction to measure the amount of protein present. The BCA kit provides a simple, reliable, and sensitive method for quantifying protein levels.
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2 protocols using bicinchoninic acid protein assay kit bca kit
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Quantifying Protein Concentration via BCA Assay
2
Characterization of Polysaccharide Structure
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Total uronic acid content was determined by a colorimetrical method [17 (link)] using glucuronic acid as a standard. Protein content was measured by a Bicinchoninic Acid Protein Assay Kit (BCA kit) (Sigma-Aldrich, 3050 Spruce Street, St Louis, MO 63103, USA) [18 (link)] using bovine serum albumin as a standard. Sulfate content was assayed using an ion chromatography method [19 (link)]. Molecular weight (Mw) was determined by a high-performance liquid chromatography, coupled with a refractive index detector (Agilent Technologies, Wilmington, DE, USA), with a column of TSKgel G3000PWXL (TOSOH, Tokyo, Japan). Aqueous Na2SO4 solution (0.1 mol/L) was used as the mobile phase and the flow rate was 0.5 mL/min. The temperature of the column was maintained at 35 °C. Dextrans were used as standards to calibrate the column [20 (link)].
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