The assay was run automated as the absorbance change at 100 seconds and 400 seconds. Tools and reagents for improved quantitation of total or specific proteins. Us4839295a measurement of protein using bicinchoninic. Piercea protein assay kit bc thermo fisher scientific. Comparisons to a minibca assay workflow indicate that insitu analysis is more accurate. The absorbance at 562 nm of the cuibicinchoninate2 complex is directly proportional to protein concentration. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from nonionic detergents and buffer salts. Although such methods are of relatively recent origin, a considerable and rapidly increasing number are already available for the assay of many of the amino acids in purified proteins and natural products. 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 bca protein assay is used for quantitation of total protein in a sample. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. Tools and reagents for improved quantitation of total or. Using the micro bca protein assay kit pierce company. Improvements in sensitivity are evident relative to standard protocols using 20.
After clearing the lysates by centrifugation at 16,000. Qubit, bradford or bicinchoninic acid in related conceptions. Activates mglu 4 receptors in transiently transfected hek293 cells. Insitu microvolume bicinchoninic acid protein assay. Although bicinchoninic acid bca has been widely used to determine protein concentration, the mechanism of interaction between protein, copper ion and bca in this assay is still not well known. Best bicinchoninic acid bca protein quantification. This protocol describes a method of quantifying protein that is a variation of the lowry assay. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. The bicinchoninic acid bca assay, first described by smith et al. The bicinchoninic acid method is faster and easier than lowry, with. The color produced is stable and increases in a linear fashion over a broad working range of increasing protein concentration. Bca protein assay reagent bicinchoninic acid youtube. Protein determination is one of the most common operations performed in biochemical research. Download msds pdf search msds currently available for download bioworld offers bca bicinchoninic acid protein assay kit with bsa standards for your research at low price.
Bicinchoninic acid protein assay and bicinchoninic acid protein assay micro ohkits are formulations based on bicinchoninic acid for the rapid and sensitive detection and quantitation of total protein content. When it is necessary to determine the total protein concentration in a. This assay is based on the protocol described by bernfeld 1951. The intracellular total ros activity assay kit red provides a sensitive, onestep fluorimetric assay to detect intracellular ros in live cells with 1 hour incubation. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from. Callan from the lankenau hospital research institute, philadelphia received for publication, june 17, 1938 the best method at present available for the exact titrimetric. The assay deviates from linearity at analyte concentrations beyond this range. Quantumprotein bicinchoninic acid protein assay1,2,3 kit is formulated for the colorimetric detection and quantitation of total protein. Single amino acids and dipeptides do not give the biuret reaction, but. The isocratic method was used together with a reversephase c18 column, acetonitrile mobile phase and water acidified with orthophosphoric acid, and a 7 min run time, according to the parameters. Improved versions of this method 2 are useful in the range of 1 to 20 nmol sugar and have been applied to the assay of endoglucanase activity 5. Insitu microvolume bicinchoninic acid protein assay peter brescia, msc. Development and validation of a hplc analytical assay. Determination of protein covalently bound to agarose supports.
Pierce bca protein assay kit thermo fisher scientific. Bicinchoninic acid bca is one of the most popular reagents for colorimetric protein determinations. The amount of reduction is proportional to the protein present. The present study describes the development and validation of an analytical assay method for efavirenz raw material and tablets by hpcl. The detergentcompatible method provides for accurate determination of protein. The bicinchoninic acid bca assay is available in kit form from pierce rockford, ill.
Bca is less variable and less susceptible to detergents than bradford, while bradford assay has a simpler and faster procedure. Bca is a highly sensitive and selective detection reagent for the. Estimation of peptide concentration by a modified bicinchoninic acid assay article in analytical biochemistry 3931. The two assays are of similar sensitivity, but since bca is stable under alkali conditions. 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. Since then it has become the most popular method for colorimetric detection and quantitation of total protein. The a 600nm response was found to be linear at least over the range 0. Bicinchoninic acid protein assay kit bca1 technical. Introduction the modulus by turner biosystems and the bca protein assay kit from pierce provides a simple method for quantifying protein over a large range. Bca protein assay reagent bicinchoninic acid from thermo. 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. The response of the bca chromophore to various interfering substances has provided insight into the mechanism of protein quantitation by.
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. The bca protein assay was introduced by smith, et al. A stock bca solution contains the following ingredients in a highly alkaline solution with a ph 11. The purple colored cubicinchoninic acid2 3 complex displays a strong absorbance at 562 nm which is proportional to protein concentration over a broad. The kit is an optimized mix and read assay format that is compatible with hts liquid handling instruments. Decreases camp levels in native cultured cerebellar granule cells. The thermo scientific pierce bca protein assay remains one of the most popular protein assay methods worldwide. Although the mechanism of color formation for the lowry assay is similar to that of. The method is relatively easy to perform, reliable, and. Bicinchoninic acid bca method an example of an indirect. The bca is used for the same reasons the lowry is used. Bicinchoninic acid assay science exchange lets you compare quotes from over 10 leading service providers.
Rapid colorimetric assays to qualitatively distinguish rna. Modified bicinchoninic acid assay for accurate determination of. Bicinchoninic acid assay an overview sciencedirect topics. Stoscheck 1990 has suggested that the bca assay will replace the lowry because it requires a single step, and the color reagent is. Most colorimetric protein assay methods can be divided into two groups based on the. Bicinchoninic acid protein assay kit is a formulation based on bicinchoninic acid for the rapid and sensitive detection and quantitation of total protein content.
Bicinchoninic acid bca assay in low volume sciencedirect. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this. A new protein assay may be adequate in different experiments beyond or instead of the wellknown standard protocols e. Subtract the average absorbance of the blank standard from the samples and. Material safety data sheet 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. Thermo scientific pierce protein assay technical handbook. Competitive binding to cuprous ions of protein and bca in the bicinchoninic acid protein assay tao huang, mian long, bo huo center for biomechanics and bioengineering and key laboratory of microgravity national microgravity laboratory, institute of mechanics, chinese academy of sciences, beijing 100190, peoples republic of china. Competitive binding to cuprous ions of protein and bca in. 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. An adaptation of pierces bca protein assay reagent kit product no. Providers we are changing the way providers access and engage customers to streamline the sale and.
Pierce chemical technical library 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. View product specific information, msds, references and buying faq. The bicinchoninic acid bca 2 assay is a colorimetric method that is commonly employed to estimate the concentration of protein in a sample. Accurately measure the total protein concentration in your sample with this twocomponent, highprecision, detergentcompatible assay reagent set. This microbiological method is used to quantitatively measure serum and red blood cell rbc folate. Also counteracts excitotoxic neuronal cell death induced by nmda in mixed cultures of cortical cells. This definition appears frequently and is found in the following acronym finder categories.
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. This procedure is very applicable to microtiter plate methods. We determined the protein concentration by the bicinchoninic acid assay sigma, using bovine serum albumin as. One particular benefit is that, unlike other methods available at that time e. Chemistry of protein assays thermo fisher scientific us. Bicinchoninic acid bca protein assay, gbiosciences vwr. Since bca is stable, it is incorporated in the reagent formulation at. Unfortunately this type of reductometric assay is not speci. 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 plot of absorbance versus analyte began to plateau. 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.
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