Subtract the average absorbance of the blank standard from the samples and. Bicinchoninic acid protein assay kit bca1 technical bulletin. Bicinchoninic acid is a highly specific chromagenic reagent for cu i forming a complex with an absorbance maximum at 562 nm. Qconcat quantification can be determined by a colorimetric protein assay such as the bicinchoninic acid assay. This procedure is very applicable to microtiter plate methods. More sensitive than lowry and less subject to interferences. The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein.
The reaction results in the development of an intense purple color with an absorbance maximum at 562 nm. The most common protein assay reagents involve either proteindye binding chemistry coomassiebradford or proteincopper chelation chemistry biuretbca. The thermo scientific pierce bca protein assay kit is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Quantumprotein bicinchoninic acid protein assay1,2,3 kit is formulated for the colorimetric detection and quantitation of total protein. Bicinchoninic acid bca protein assay photometric method reading at 562 nm. Tools and reagents for improved quantitation of total or. Box 117 industriezone iii rockford, il 61105 industrielaan 27 usa 9320 erembodegemaalst 815.
Otherwise, contact supplier for specific applications. Pierce bca protein assay kit brigham young university. Pdf bicinchoninic acid bca assay in low volume researchgate. The protein protocols handbook, second edition aims to provide a crosssection of analytical techniques commonly used for proteins and peptides, thus providing a benchtop manual and guide for those who are new to the protein chemistry laboratory and for those more established workers who wish to use a technique for the first time. Sulfonhydroxysuccinimide interferes with bicinchoninic acid protein assay. In 96well plates, the relationship between protein content and. Pdf measurement of protein using bicinchoninic acid m. The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex. Bicinchoninic acid assay an overview sciencedirect topics.
The purple colored cubicinchoninic acid2 3 complex displays a strong absorbance at 562 nm which is proportional to protein concentration over a broad. Protein determination by the bicinchoninic acid bca method. The bca protein assay is used for quantitation of total protein in a sample. Rapid 2,2bicinchoninicbased xylanase assay compatible with. Jeffries institute for microbial and biochemical technology, usda forest service, forest products laboratory, one gifford pinchotdrive, madison, wi 537262398,usa. Although these methods are highly accurate when used to assay. The same pure protein should be used as a standard for the most accurate results. Piercea protein assay kit bc thermo fisher scientific. A new protein assay may be adequate in different experiments beyond or instead of the wellknown standard protocols e. Bicinchoninic acid protein assay and bicinchoninic acid protein assay micro kits are formulations based on bicinchoninic acid for the rapid and sensitive detection and quantitation of total protein content. Protein quantitation based on bicinchoninic acid bca is simple, sensitive, and tolerant to many detergents and substances known to interfere with the lowry method. Pdf the bca assay is a colorimetric method for estimating protein concentration.
Rapid 2,2bicinchoninicbased xylanase assay compatible with high throughput screening william r. In particular, it is insensitive to detergents such as triton x100 and sds 1 %. Interferences attributed to glucose, ascorbic acid, and uric acid in this reaction were significantly reduced by performing a twopoint assay and by including borate ions in the buffer. Us4839295a measurement of protein using bicinchoninic acid. Qubit, bradford or bicinchoninic acid in related conceptions.
Chemistry of protein assays thermo fisher scientific za. The bicinchoninic acid bca assay is available in kit form from pierce rockford, ill. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from. Linear working range for bsa is 20 2000 gml for the bicinchoninic acid protein assay. Pdf measurement of protein using bicinchoninic acid m n. Expired lifetime application number us07053,708 inventor paul k. Bca protein assay kit ab102536 provides a simple, rapid, detergent tolerant up to 5% procedure for determining the concentration of proteins in solution. First is the biuret reaction, whose faint blue color results from the reduction of cupric ion to cuprous ion. The bicinchoninic acid bca protein assay primarily relies on two reactions. Bca product description protein determination is one of the most common operations performed in biochemical research. Bca bicinchoninic acid protein assay fanglian he department of.
The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this assay has the advantage that it can be. These assays offer low protein consumption and convenience, but variable accuracy depending on the amino acid composition of the qconcat protein. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. Unlike the bradford and the lowry, the bca assay features a relatively small proteintoprotein variation 11, 28, making it the most suitable assay for total protein quantification. The bicinchoninic acid bca assay, first described by smith et al.
Bicinchoninic acid protein assay kit catalog numbers bca1 and b9643 technical bulletin synonym. Measurement of protein using bicinchoninic acid sciencedirect. The micro bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions and is a modification of the bicinchoninic acid bca protein assay for dilute protein samples 0. The bicinchoninic acid bca assay for protein quantitation. Of these, the test tube procedure requires a larger volume 0. The absorbance at 562 nm of the cuibicinchoninate2 complex is directly proportional to protein concentration. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. This assay is carried out in a volume of 1 ml, using 50. Bicinchoninic acid protein assay kit bca1 technical. Sulfonhydroxysuccinimide interferes with bicinchoninic acid. Bicinchoninic acid bca protein assay smith rice university.
Bicinchoninic acid bca protein assay, gbiosciences vwr. The bicinchoninic acid assay bca, also referred to as the smiths assay, is a biochemical assay designed by paul smith in 1985. Quantifying protein by bicinchoninic acid csh protocols. Jun 17, 2016 in the bca assay, the folinciocalteu reagent is replaced with bicinchoninic acid as described by smith. The thermo scientific pierce bca protein assay kit is a. Abstract the bca protein assay is used for quantitation of total protein in a sample. The bca is used for the same reasons the lowry is used.
Because of this property, the resultant absorbance at 562 nm is directly proportional to the protein concentration. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric. Reagents bicinchoninic acid solution, catalog number b9643 reagent a is a 1,000 ml solution containing bicinchoninic acid, sodium carbonate, sodium tartrate, and sodium bicarbonate in 0. Rapid solidphase determination of cereal protein using. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutionsaccurately quantifying total protein concentration is a key to most experiments and is often necessary for isolation, separation, and analysis of proteins by biochemical methods. The two assays are of similar sensitivity, but since bca is stable under alkali conditions. Smith at the pierce chemical company, now part of thermo fisher scientific, is a biochemical assay for determining the total concentration of protein in a solution 0. Synonyms bicinchoninic acid protein assay reagent a intended use refer to the instruction booklet for proper and intended use. Pierce bca protein assay kit thermo fisher scientific.
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