Pierce bca protein assay kit brigham young university. Chemistry of protein assays thermo fisher scientific za. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this assay has the advantage that it can be. Bicinchoninic acid protein assay kit bca1 technical.
Bicinchoninic acid bca protein assay, gbiosciences vwr. Some of the variables and the interferences which affect the reaction kinetics of proteins with bicinchoninic acid bca 4,4dicarboxy2,2biquinoline have been studied. Rapid 2,2bicinchoninicbased xylanase assay compatible with high throughput screening william r. The bicinchoninic acid bca protein assay primarily relies on two reactions. Synonyms bicinchoninic acid protein assay reagent a intended use refer to the instruction booklet for proper and intended use. Jeffries institute for microbial and biochemical technology, usda forest service, forest products laboratory, one gifford pinchotdrive, madison, wi 537262398,usa. Measurement of protein using bicinchoninic acid sciencedirect. Sulfonhydroxysuccinimide interferes with bicinchoninic acid. The bca protein assay is used for quantitation of total protein in a sample. The thermo scientific pierce bca protein assay kit is a. Pdf the bca assay is a colorimetric method for estimating protein concentration. Qubit, bradford or bicinchoninic acid in related conceptions. Jun 17, 2016 in the bca assay, the folinciocalteu reagent is replaced with bicinchoninic acid as described by smith.
Bicinchoninic acid protein assay kit bca1 technical bulletin. Pierce bca protein assay kit thermo fisher scientific. The purple colored cubicinchoninic acid2 3 complex displays a strong absorbance at 562 nm which is proportional to protein concentration over a broad. 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. Bicinchoninic acid assay an overview sciencedirect topics. Bicinchoninic acid bca protein assay photometric method reading at 562 nm. These assays offer low protein consumption and convenience, but variable accuracy depending on the amino acid composition of the qconcat protein. The two assays are of similar sensitivity, but since bca is stable under alkali conditions. More sensitive than lowry and less subject to interferences. Bca bicinchoninic acid protein assay fanglian he department of. 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.
Bicinchoninic acid protein assay kit catalog numbers bca1 and b9643 technical bulletin synonym. 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. Subtract the average absorbance of the blank standard from the samples and. 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. Rapid solidphase determination of cereal protein using. Pdf measurement of protein using bicinchoninic acid m n. In particular, it is insensitive to detergents such as triton x100 and sds 1 %. 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.
The reaction results in the development of an intense purple color with an absorbance maximum at 562 nm. The bca is used for the same reasons the lowry is used. Otherwise, contact supplier for specific applications. Linear working range for bsa is 20 2000 gml for the bicinchoninic acid protein assay. A new protein assay may be adequate in different experiments beyond or instead of the wellknown standard protocols e. This procedure is very applicable to microtiter plate methods. The bicinchoninic acid assay bca, also referred to as the smiths assay, is a biochemical assay designed by paul smith in 1985.
Because of this property, the resultant absorbance at 562 nm is directly proportional to the protein concentration. Quantifying protein by bicinchoninic acid csh protocols. Bca product description protein determination is one of the most common operations performed in biochemical research. The same pure protein should be used as a standard for the most accurate results. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed. Sulfonhydroxysuccinimide interferes with bicinchoninic acid protein assay. The bicinchoninic acid bca assay is available in kit form from pierce rockford, ill. Expired lifetime application number us07053,708 inventor paul k. 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.
Abstract the bca protein assay is used for quantitation of total protein in a sample. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. The absorbance at 562 nm of the cuibicinchoninate2 complex is directly proportional to protein concentration. The bicinchoninic acid bca assay for protein quantitation. This assay is carried out in a volume of 1 ml, using 50. The bicinchoninic acid bca assay, first described by smith et al. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric. Bca protein assay kit ab102536 provides a simple, rapid, detergent tolerant up to 5% procedure for determining the concentration of proteins in solution. Qconcat quantification can be determined by a colorimetric protein assay such as the bicinchoninic acid assay. Bicinchoninic acid bca protein assay smith rice university. Protein quantitation based on bicinchoninic acid bca is simple, sensitive, and tolerant to many detergents and substances known to interfere with the lowry method. Of these, the test tube procedure requires a larger volume 0.
The most common protein assay reagents involve either proteindye binding chemistry coomassiebradford or proteincopper chelation chemistry biuretbca. Although these methods are highly accurate when used to assay. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. Box 117 industriezone iii rockford, il 61105 industrielaan 27 usa 9320 erembodegemaalst 815. Pdf bicinchoninic acid bca assay in low volume researchgate. The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex. Protein determination by the bicinchoninic acid bca method. 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. Quantumprotein bicinchoninic acid protein assay1,2,3 kit is formulated for the colorimetric detection and quantitation of total protein. Piercea protein assay kit bc thermo fisher scientific. First is the biuret reaction, whose faint blue color results from the reduction of cupric ion to cuprous ion.
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. In 96well plates, the relationship between protein content and. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from. The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. 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. Pdf measurement of protein using bicinchoninic acid m. Bicinchoninic acid bca assay in low volume article pdf available in analytical biochemistry 4102. Tools and reagents for improved quantitation of total or.
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