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SKU AR0146
Pack size 1 kit
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays
Price: $60.00
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Overview

Product Name BCA Protein Assay Kit
Synonyms Bicinchoninic acid protein assay kit; BCA Protein Quantification Kit, BCA Protein Quantitation Kit, BCA Protein Estimation Kit, Bicinchoninic acid Protein Quantification Kit, Bicinchoninic acid Protein Quantitation Kit, Bicinchoninic acid Protein Estimation Kit, BCA kit for protein determination, Bicinchoninic acid kit for protein determination, Smith Protein Assay kit
Description BCA Protein Assay Kit is a ready-to-use detergent-compatible Western blot related total protein analysis reagent used for the quick determination of total protein concentration by measuring absorbance at 562 nm and comparing to a protein standard absorption vs. concentration curve, according to Smith.
Application Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays
Pack Size 1 kit
Reagent Type Protein assay kit; Western Blotting related
Usage Studying protein-protein interactions, Measuring column fractions after affinity chromatography, Assessing protein yields in whole cell lysates, High-throughput screening of fusion proteins

Assay Information

Assay Type Protein Quantitative Measurement
Sample Type Serum, Plasma, Cell culture extracts, Tissue Extracts
Technique Solution-based detection of spectral absorbance at 562 nm; Smith Protein assay
Detection Method Colorimetric/Spectroscopic
Equipment needed Тest tubes or miscroplates; Spectrophotometer; Microplate Reader
Assay Purpose Measure protein concentration in solution
Assay Range 20 - 2000 μg/ml
Incubation Requirements 30 min at 37°C
Kit components BCA Reagent A: 100 ml
BCA Reagent B: 5 ml
Albumin (BSA) Standards: 20 ml (2 mg/ml)

Properties

Form Supplied BCA Reagent A and B: ready-to-use 1X solutions
BSA Standard: stock solution for preparing a series of standard protein dilutions
Physical State of A+B Reagent mixture Light blue liquid
Recommended working concentration See protocol in Datasheet
Reagent Color/Absorbance Blue / A562 nm
Activity Active in alkaline medium
Assays per kit Tube procedure: 50 assays
Microplate procedure: 500 assays
Compatibility with reagents Compatible with typical concentrations of most ionic and nonionic detergents
Incompatible with chelating agents, strong acids or bases, and reducing agents: interfere with the reduction and chelating reactions of the assay mechanism
Storage Store at room temperature for one year.
Precautions FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE

Usage and Handling

To be used with: Lysates or homogenates, prepared with non-reducing lysis and extraction buffers: Mammal Tissue Protein Extraction Reagent, Mammal Cell Protein Extraction Reagent, RIPA Lysis Buffer, Mitochondria Extraction Reagent, Membrane Protein Extraction Kit, Cytoplasmic and Nuclear Extraction Kit, Bacterial Cell Protein Extraction Reagent

Background

The BCA Protein Assay is a detergent - compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantization of total protein. This method combines the reduction of Cu 2+ to Cu 1+ by protein in an alkaline medium (Biuret reaction) with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu 1+) by bicinchoninic acid, forming a purple-colored BCA/copper complex with absorbance maximum at 562 nm. Measurement of A562 is used to determine the concentration of protein in solutions. Protein concentrations are estimated by reference to absorbances obtained for a series of standard protein dilutions (e.g. of BSA), which are assayed alongside the unknown sample. The BCA assay involves two consecutive steps of color development, where the final product indirectly depends on the protein concentration in the solution. The first step is the chelation of copper with protein in an alkaline environment, followed by reduction of the Cu2+ (cupric) to Cu1+ (cuprous) cations forming a light blue complex. It has been shown that cysteine, cystine, tryptophan, tyrosine, and the peptide bond are able to reduce Cu2+ to Cu1+. The amount of reduction is proportional to the amount of protein present (the number of peptide bonds). In the second step of the color development reaction, bicinchoninic acid (BCA), a highly specific chromogenic reagent for Cu1+, reacts with the reduced cuprous cation that was formed in step one, producing a purple complex with strong absorbance at 562 nm (Smith 1985, Wiechelman 1988). The purple-colored reaction product results from the chelation of two molecules of BCA with one cuprous ion. The intensity of the BCA/copper purple complex is proportional to the amount of protein in the sample. Thus measuring the intensity of the absorbance at 562 nm is proportional to protein concentration. The BCA protein assay demonstrates higher tolerances towards common interfering substances, such as nonionic detergents and buffer salts, than the Lowry technique.

BCA Protein Assay Kit Images

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BCA Protein Assay Kit
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Publications

Xiong, Q., Ru, Q., Chen, L., Tian, X., & Li, C. (2017). Mitochondrial dysfunction and inflammatory response in the cytotoxicity of NR8383 macrophages induced by fine particulate matter. Environmental Toxicology and Pharmacology, 55, 1-7. Advance online publication. doi: 10.1016/j.etap.2017.07.017
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FAQs

Q: What is the correct storage temperature of this kit? On the manual it says room temperature, but in the manual that we received in the past it said 4 degrees.
A: We have modified the storage temperature of this kit as room temperature recently, but it can be stored at 4 degree as well. The BSA solution contains sodium azide, it is available at room temperature.