The bicinchoninic acid bca assay is available in kit form from pierce rockford, ill. The thermo scientific pierce bca protein assay kit is a detergent compatible formulation based on bicinchoninic acid bca for the. Bicinchoninic acid kit and bovine serum albumin bsa were obtained from sigmaaldrich. Pdf competitive binding to cuprous ions of protein and bca. Total protein quantitation using the bicinchoninic acid assay. Expired lifetime application number us07053,708 inventor paul k.
Pdf bicinchoninic acid bca assay in low volume researchgate. Using the matrix platemate 2x2 to automate the bca protein assay. 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. Bca bicinchoninic acid protein assay kit with bsa standards. With two commercially prepared flours microcrisp the rapid determination of protein in cerealbased materials is described. 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 is a highly specific chromagenic reagent for cu i forming a complex with an absorbance maximum at 562 nm. Bicinchoninic acid assay an overview sciencedirect topics. Pdf measurement of protein using bicinchoninic acid m n. Showing your workcalculations can be achieved by typing it out, pasting in a clear image of your handwritten work, or using handwritten annotation on the. The loosely and firmly adsorbed amount of bovine serum albumin bsa on dental titanium and dental gold was investigated by bicinchoninic acid assay bca assay.
The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to. Insitu microvolume bicinchoninic acid protein assay. The bicinchoninic acid bca assay, first described by smith et al. This procedure is very applicable to microtiter plate methods.
To overcome this limitation, we performed the bca assays in opa. Pdf bicinchoninic acid assay protein quantification. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed. Bicinchoninic acid bca assay or smith assay is a copperbased colorimetric assay for total protein quantification. 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.
Determination of protein covalently bound to agarose supports. The bicinchoninic acid bca assay for protein quantitation methods mol biol. Characterization of protein films on dental materials. More sensitive than lowry and less subject to interferences. Total protein quantitation using the bicinchoninic acid. Us4839295a measurement of protein using bicinchoninic acid.
Bicinchoninic acid protein assay kit bca1 technical. Since bca is stable, it is incorporated in the reagent formulation at. Piercea protein assay kit bc thermo fisher scientific. In 96well plates, the relationship between protein content. Smith at the pierce chemical company in 1985 smith et al. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. Briefly the bca assay comprised mixing 10 l bca working reagent with 10 l. Tools and reagents for improved quantitation of total or. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric. The bca is used for the same reasons the lowry is used. The most common protein assay reagents involve either proteindye binding chemistry coomassiebradford or proteincopper chelation chemistry biuretbca.
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 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. Authors anthony bainor 1, lyra chang, thomas j mcquade, brian webb, jason e gestwicki. We investigated the ability of gradient elution moving boundary electrophoresis gembe with capacitively coupled contactless conductivity detection c 4 d to assay total protein concentration using the bicinchoninic acid bca reaction. The bicinchoninic acid bca protein assay is highly sensitive colorimetric assay that is not affected by chemicals in the sample.
Rapid 2,2 bicinchoninic based xylanase assay compatible with high. In 96well plates, the relationship between protein content and absorbance is nearly linear over a wide range. Pierce bca protein assay kit university of arizona. Some of the variables and the interferences which affect the reaction kinetics of proteins with bicinchoninic acid bca 4,4dicarboxy2,2biquinoline have been studied. This spectrophotometric assay is used to determine the enhancement of adriamycin cytotoxicity against resistant cancer cells by plant extracts or pure compounds indirectly. The bca assay is a colorimetric method for estimating protein concentration. Rapid solidphase determination of cereal protein using.
Pdf the bca assay is a colorimetric method for estimating protein concentration. Bicinchoninic acid bca assay in low volume anal biochem. The color produced is stable and increases in a linear fashion over a broad working range of increasing protein concentration. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. Protein assay using bicinchoninic acid 81 lent standard curves, but with variable slopes. Synonyms bicinchoninic acid protein assay reagent a intended use refer to the instruction booklet for proper and intended use. 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.
The bca protein assay is used for quantitation of total protein in a sample. Accurately measure the total protein concentration in your sample with thi. Stoscheck 1990 has suggested that the bca assay will replace the lowry because it requires a single step, and the color reagent is. Pdf bicinchoninic acid assay find, read and cite all the research you need on researchgate. Rapid 2,2bicinchoninicbased xylanase assay compatible. Detection and quantitative estimation of proteins by. Protein determination by the bicinchoninic acid bca method. The bicinchoninic acid assay bca, also referred to as the smiths assay, is a biochemical assay designed by paul smith in 1985. 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. In bca the color develops in a single step but lowry method in two steps. Bca protein assay reagent bicinchoninic acid youtube.
Rapid 2,2bicinchoninicbased xylanase assay compatible with. Bca offers the advantage of being highly sensitive and specific for cui which rapidly forms an intense purple complex in the presence of bca. The bicinchoninic acid bca assay for protein quantitation. The two assays are of similar sensitivity, but since bca is stable under alkali conditions.
A variety of temperatures and ph conditions can be used and, after incubation, the assay requires only one detection reagentand one heating step. Affiliation 1 department of pathology and the life. Protein determination by the bicinchoninic acid bca. Chemical product and company identification responsible namepierce administration synonyms bicinchoninic acid. Abstract the bca protein assay is used for quantitation of total protein in a sample. This method does not require special properties of the sample like a high reflectivity or conductivity and turns the bca assay into a valuable tool to investigate a widespread spectrum. The bicinchoninic acid bca assay is a protein quantification technique that was first demonstrated by paul k. The bca assay was demonstrated using two microvolume formats. The protein contents were determined using the bicinchoninic acid assay bca 43 and a calibration curve built with standard bovine serum albumin.
Pdf bicinchoninic acid assay protein quantification technical. Our improved formulation, the bc assay, provides significant benefits over existing colorimetric assays, namely sensitivity, reagent stability, and detergent compatibility. Pierce bca protein assay kit thermo fisher scientific. The reaction results in the development of an intense purple color with an absorbance maximum at 562 nm. Us4839295a measurement of protein using bicinchoninic. No one reagent can be considered to be the ideal or best protein assay method. Us5763282a absorbance assay for the measurement of. Subtract the average absorbance of the blank standard from the samples and. The bicinchoninic acid protein assay has been adapted for use in a microtiter plate, into an easy, indirect method for screening mdr efflux blockers in plant extracts. Doc bicinchoninic acid bcabased protein assays chemistry. Please denote your answers by using bold, italic s, or colored font.
Author j m walker 1 affiliation 1 division of biosciences, university of hertfordshire, hatfield, uk. The choice among available protein assays is usually based on the compatibility of the protein assay method with the samples. Bicinchoninic acid bca protein assay panreac applichem itw. The absorbance at 562 nm of the cuibicinchoninate2. A quantitative test for copper using bicinchoninic acid. Competitive binding to cuprous ions of protein and bca in the bicinchoninic acid protein assay. Instead, bicinchoninic acid based assay was, indeed, distinctly affected by the presence of nanoparticleiron in suspension 0. Thermo scientific pierce protein assay technical handbook. The first format was the minibca assay recommended by the nanodrop technical manual 2. 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. This protocol describes a method of quantifying protein that is a variation of the lowry assay. N2 we describe a direct colorimetric assay for copper in serum and biological samples using 2,2. Bicinchoninic acid protein assay in the determination of. The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex.
Box 117 industriezone iii rockford, il 61105 industrielaan 27. A rapid, practical assay for measuring the concentration of cornified envelopes based on measuring the absorbance, which utilizes the addition of bicinchoninic acid to insoluble cornified envelopes preparation, after the cornified envelopes have been separated from soluble proteins. The bicinchoninic acid bca protein assay primarily relies on two reactions. Bca protein assay reagent a, 500 ml 3971 0023228 call chemtrec. Adaptation of the bicinchoninic acid protein assay for use with microtiter plates and sucrose gradient fractions. The assay is linear for sugar concentrations from 0 to 86 gml. Because of this property, the resultant absorbance at 562 nm is directly proportional to the protein concentration. The purple colored cu bicinchoninic acid 2 3 complex displays a strong absorbance at 562 nm which is proportional to protein concentration over a broad.
Bca bicinchoninic acid protein assay fanglian he department of. A modification of the bicinchoninic acid assay that can be used for between replicates. The principle of this technique is quite similar to the lowry protein assay, which is one of the alternative protein quantification methods. C 4 d behaves as a concentration dependent detection system, unlike optical methods that also rely on pathlength due to. The bicinchoninic acid protein assay kit for protein quantitation and determination is formulated for. Pdf measurement of protein using bicinchoninic acid m. Using the matrix platemate 2x2 to automate the bca protein.
Bicinchoninic acid protein assay kit bca1 technical bulletin. Otherwise, contact supplier for specific applications. Bca bicinchoninic acid protein assay is watersoluble and strongly absorbs light at 562nm in a linear fashion with increasing protein concentration. 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. The bca protein assay offers an accurate measurement of protein concentration for a large linear range 202,000 gml and is compatible with a wide variety of buffers and solvents2. Bicinchoninic acid protein assay kit is a formulation based on bicinchoninic acid for the rapid and sensitive detection and quantitation of total protein content. Each method has its advantages and disadvantages table 1. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from nonionic detergents and buffer salts. The amount of reduction is proportional to the amount of protein present.
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