Bicinchoninic acid bca protein assay, gbiosciences vwr. Bicinchoninic acid bca reducing agent compatible protein assay. Tools and reagents for improved quantitation of total or. Storage and stability 12 months at room temperature introduction quantumprotein bicinchoninic acid protein assay1,2,3 kit is formulated for the colorimetric detection and quantitation of total protein. The bca assay is a colorimetric method for estimating protein concentration. Bicinchoninic acid bca reducing agent compatible protein. Pdf bicinchoninic acid assay protein quantification. 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. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. 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. Thermo scientific pierce protein assay technical handbook. The method is based on reaction with a reagent consisting of acetic anhydride and concentrated sulfuric acid. Bicinchoninic acid kit for protein determination bca.
To overcome this limitation, we performed the bca assays in opaque, white 384well plates. Ascorbic acid and uric acid tyrosine, cysteine and tryptophan 50mm imidazole, 0. The bca protein assay was introduced by smith, et al. Best bicinchoninic acid bca protein quantification. Article information, pdf download for a rapid transglutaminase assay for high throughput. A novel sandwich elisa for the quantitative and sensitive determination of il in human serum and plasma was established. Those peptide groups are then free to bind another molecule of cupric ion. Bicinchoninic acid is most commonly employed by biochemists in the bicinchoninic acid bca assay, which is used to determine the total level of protein in a solution. Since then it has become the most popular method for colorimetric detection and quantitation of total protein. The two assays are of similar sensitivity, but since bca is stable under alkali conditions. Acid base chemistry notes pdf 10p this note covers the following topics. The first format was the minibca assay recommended by the nanodrop technical manual 2.
Blank titrations in acetic acid and the effect of added water. These assays offer low protein consumption and convenience, but variable accuracy depending on the amino acid composition of the qconcat protein. Begley a, a department of chemistry and chemical biology, cornell university, ithaca, ny 14853, usa b department of natural resources, cornell university, ithaca, ny 14853, usa received 8 november 2006 available online 7 june 2007 abstract an alternative method for. Bronsted lowry acid, lewis acid, acids in aqueous solution, relationship between kb and ka, muliprotic acids, equilibrium calculations, mass balance, equilibrium relationship, proton condition, weak acidbase calculations, strong acidbase calculations, strong acidstrong base mixtures, strong acidweak base mixtures. Adaptation of the bicinchoninic acid protein assay for use with microtiter plates and sucrose gradient fractions. The same pure protein should be used as a standard for the most accurate results. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from. Protocol overview the jejunum of chickens are fragmented and resuspended in lysis buffer 1x. For more results try searching for bicinchoninic acid assay across all experimental services.
Discussions focus on serial dilution assay, requirements for accurate assay, microbiological assay of riboflavin, laws of adsorption and partition, mechanisms of. Sample volume per 200300l total volume for measurement in 96well microplate. The assay was validated and was shown to be accurate and precise over the entire quantification range. Use a protein assay that is unaffected by interfering agents ni. Sialic acid assay kit dsla100 bioassay systems 877. 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. Download bca bicinchoninic acid protein assay kit with bsa standards msds pdf. The bca assay was demonstrated using two microvolume formats. Determination of protein concentration using bicinchoninic acid bca protein assay kit.
Bca protein assay protocol a solutions and reagents kit contents. All samples must be free of organic solvents prior to assay. Sialic acid is oxidized to formylpyruvic acid which reacts with thiobarbituric acid. A spectrophotometric method of analysis of fusidic acid. The most common protein assay reagents involve either proteindye binding chemistry coomassiebradford or proteincopper chelation chemistry biuretbca. Bicinchoninic acid protein assay kit bca1 technical bulletin. Instead, bicinchoninic acid based assay was, indeed, distinctly affected by the presence of nanoparticleiron in suspension 0. When it is necessary to determine the total protein concentration in a. Synonyms bicinchoninic acid protein assay reagent a intended use refer to the instruction booklet for proper and intended use.
The use of raca allows for samples containing up to 5mm dtt, 10mm tcep or 35mm. Qubit, bradford or bicinchoninic acid in related conceptions. All multiplex assays present significant challenges, and laboratory personnel can develop assays inhouse or use commercially available multiplex assays involving a variety of. Box 117 industriezone iii rockford, il 61105 industrielaan 27 usa 9320 erembodegemaalst 815. Jul 23, 2014 bca bicinchoninic acid protein assay the bca assay is based on the traditional lowry assay except bicinchoninic acid is used resulting in a very pretty purple color that absorbs at 560 nm. In principle, a solution containing a free aldohexose such as glucose, liberated from a glucosylated protein via acid hydrolysis, would test positive by the benedicts assay. Decreases camp levels in native cultured cerebellar granule cells. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from nonionic detergents and buffer salts. The color produced is stable and increases in a linear fashion over a broad working range of increasing protein concentration. Competitive binding to cuprous ions of protein and bca in. Qconcat quantification can be determined by a colorimetric protein assay such as the bicinchoninic acid assay. The assay was validated and was shown to be accurate and precise over the entire quantification range 0.
Quantummicroprotein bicinchoninic acid protein assay1,2,3 kit is formulated for the colorimetric detection and quantitation of total protein in dilute samples 0,5 20 gml. Sample volume per 1ml total assay volume for measurement in 1cm cuvette standard protocol. A rapid transglutaminase assay for highthroughput screening. Bicinchoninic acid kit and bovine serum albumin bsa were obtained from sigmaaldrich. An assay for thiaminase i in complex biological samples. Total protein content was measured by a standard bicinchoninic acid assay bca assay. Bronsted lowry acid, lewis acid, acids in aqueous solution, relationship between kb and ka, muliprotic acids, equilibrium calculations, mass balance, equilibrium relationship, proton condition, weak acidbase calculations, strong acidbase calculations, strong acidstrong base mixtures, strong. Piercea protein assay kit bc thermo fisher scientific. An assay for thiaminase i in complex biological samples jeremiah w. Next, we use the bicinchoninic acid bca protein assay from g bioscience to quantify total proteins. A spectrophotometric method for assay of fusidic acid is described. All quotes are valid for 30 days unless otherwise stated. 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. Download our protein assays and bioassays handbooks.
Quantumprotein bicinchoninic acid protein assay1,2,3 kit is formulated for the colorimetric detection and quantitation of total protein. Bicinchoninic acid bca method one of the most recent advances in protein assay technology is the use of bicinchoninic acid bca in a unique reagent system. Bicinchoninic acid protein assay kit bca1 technical. The bca assay is based on the traditional lowry assay except bicinchoninic acid is used resulting in a very pretty purple color that absorbs at 560 nm. Pancreatic lipase monitoring is also used to help diagnose crohns disease, cystic fibrosis and. Tocris products are intended for laboratory research use only, unless stated otherwise. Since bca is stable, it is incorporated in the reagent formulation at. Total cholesterol and cholesteryl ester colorimetric assay kit ii.
Briefly the bca assay comprised mixing 10 l bca working reagent with 10 l. 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. Bca bicinchoninic acid protein assay kit with bsa standards. Insitu microvolume bicinchoninic acid protein assay. May 19, 2016 nucleic acid testing with multiple assays, or when two or more targets are detected simultaneously as set through a preparation process, is a complicated procedure. The bicinchoninic acid bca assay method for the determination of protein has been investigated for its utility in measuring the. The assay employs an incubation step at acidic ph, which was shown to decrease nonspecific binding and interference from il binding proteins. A new protein assay may be adequate in different experiments beyond or instead of the wellknown standard protocols e. The bicinchoninic acid bca assay for protein quantitation. However, in practice a positive result for glycoproteins. The bicinchoninic acid bca protein assay primarily relies on two reactions. This corresponds to a tartaric acid concentration of 54 mgl of sample solution at. Sialic acid assay kit dsla100 bioassay systems 877782.
For batch specific data refer to the certificate of analysis. Bca is sensitive and has a broad dynamic range capable of measuring protein concentrations of 0. Total dna content was determined by measuring the absorbance at 260 nm. Bicinchoninic acid kit protein determination is one of the most common operations performed in biochemical research. Bicinchoninic acid bca kit v0773 test optionsvariations. Otherwise, contact supplier for specific applications. Us4839295a measurement of protein using bicinchoninic. Activates mglu 4 receptors in transiently transfected hek293 cells. 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. Nucleic acid testing with multiple assays, or when two or more targets are detected simultaneously as set through a preparation process, is a complicated procedure. To evaluate the performance of each of the purification methods performed earlier, purified omvs were evaluated by several methods as summarized in table 1. The binding of bca to cuprous ion effectively removes the weakly chelated peptides of the biuret reaction. Megacalctm, downloadable from where the product appears on. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this assay has the advantage that it can be.
The bicinchoninic acid bca assay, first described by smith et al. Therefore, if bicinchoninic acid and copper are present in large excess as they always are in bca protein assay reagents, the protein assay does not reach an end. Bicinchoninic acid protein assay kit is a formulation based on bicinchoninic acid for the rapid and sensitive detection and quantitation of total protein content. The technical data provided above is for guidance only.
Experiment study of acidbase assay by conductometric method. Bca bicinchoninic acid protein assay is watersoluble and strongly absorbs light at 562nm in a linear fashion with increasing protein concentration. Bicinchoninic acid assay an overview sciencedirect topics. This protocol describes a method of quantifying protein that is a variation of the lowry assay. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this assay has the advantage that it. The smallest differentiating absorbance for the assay is 0. Rapid colorimetric assays to qualitatively distinguish rna. Also counteracts excitotoxic neuronal cell death induced by nmda in mixed cultures of cortical cells.
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