BCA Protein Colorimetric Detection Kit (Two Plate)
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|Country Availability||DK, EE, FI, IS, LT, LV, NO, SE|
|Data Sheet Link||http://www.arborassays.com/documentation/inserts/K041-H1.pdf|
|Description||BCA Protein Colorimetric Detection Kit (Two Plate)|
|Supplier||Arbor Assays Inc.|
|Notes||Protein determination is one of the most common operations performed in biochemical research. The principle of the bicinchoninic acid (BCA) assay is similar to the Lowry assay1, and relies on the formation of a Cu2+-protein complex under alkaline conditions2, followed by reduction of the Cu2+ to Cu1+. The amount of reduction is proportional to protein present. It has been shown that cysteine, cystine, tryptophan, tyrosine, and peptide bonds2 are able to reduce Cu2+ to Cu1+. BCA forms a purple-blue complex with Cu1+ in alkaline environments, thus providing a basis to monitor the reduction of alkaline Cu2+ by proteins. The method combines the well-known reduction of Cu2+ to Cu1+ by protein in an alkaline medium (the biuret reaction) with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu+1) using a unique reagent containing bicinchoninic acid. The purple-colored reaction product of this assay is formed by the chelation of two molecules of BCA with one cuprous ion. This water-soluble complex exhibits a strong absorbance at 562nm that is nearly linear with increasing protein concentrations over a broad working range (6-1,000 μg/mL). The BCA chemistry is not a true end-point method; that is, the final color continues to develop. However, following incubation, the rate of continued color development is sufficiently slow to allow large numbers of samples to be assayed together.|
|Protocol||The DetectX® BCA Protein Assay Kit is designed to quantitatively measure total protein content in a variety of samples. The assay measures all types of proteins from all species. Please read the complete kit insert before performing this assay. A bovine serum albumin (BSA) standard is provided to generate a standard curve for the assay and all samples should be read off of the standard curve. Samples are diluted in water and added to the wells. The BCA Color Solution is made by mixing the BCA Reagent with the BCA Enhancer. The BCA Color Solution is added to all wells and the plate incubated at 37°C. Protein in the samples reacts with the BCA Color Reagent to generate a purple colored product which is read at 560 nm.|
|References||1. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ., “Protein measurement with the Folin phenol reagent”. J. Biol. Chem. 1951, 193 (1): 265–75.2. Smith, PK, et. al., “Measurement of protein using bicinchoninic acid.”, Anal. Biochem., 1985, 150: (1), 76-85.|
|Sensitivity||6.65 μg/mL (Regular Format). 1.68 μg/mL (High Sensitivity Format).|
|Species Reactivity||species independent|
|Storage||All components of this kit should be stored at room temperature until the expiration date of the kit.|
|Technical Specifications||Supplied ComponentsClear 96 well Half Area Plates 2 Plates Catalog Number X018-2EACorning Costar Plate 3695. Bovine Serum Albumin Standard 200 μL Catalog Number C143-200ULBovine Serum Albumin (BSA) at 10 mg/mL BCA Reagent 16 mL Catalog Number C144-16MLBCA Reagent solution BCA Enhancer 320 μL Catalog Number C145-320ULBCA Enhancer solution containing copper sulfate Plate Sealers 2 Each Catalog Number X002-1EA|
|Product Page Updated||2019-01-03T09:12:03.477Z|
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