Rapid colorimetric assays to qualitatively distinguish rna. The amount of reduction is proportional to the protein present. One particular benefit is that, unlike other methods available at that time e. Pierce bca protein assay kit, sufficient reagents for 500 testtube or 5000 microplate. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric.
Determination of protein covalently bound to agarose supports. Estimation of peptide concentration by a modified bicinchoninic acid assay article in analytical biochemistry 3931. Activates mglu 4 receptors in transiently transfected hek293 cells. Stoscheck 1990 has suggested that the bca assay will replace the lowry because it requires a single step, and the color reagent is. Material safety data sheet bca protein assay reagent a, 500 ml 3971 0023228 call chemtrec. Also counteracts excitotoxic neuronal cell death induced by nmda in mixed cultures of cortical cells. Bicinchoninic acid protein assay kit bca1 technical. The present study describes the development and validation of an analytical assay method for efavirenz raw material and tablets by hpcl. Bca is a highly sensitive and selective detection reagent for the.
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. This assay is based on the protocol described by bernfeld 1951. Accurately measure the total protein concentration in your sample with this twocomponent, highprecision, detergentcompatible assay reagent set. Chemical product and company identification responsible namepierce administration. Most colorimetric protein assay methods can be divided into two groups based on the. Bca protein assay reagent bicinchoninic acid from thermo. The plot of absorbance versus analyte began to plateau. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this. Bca protein assay reagent bicinchoninic acid youtube.
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 microplate procedure affords the sample handling ease of a microplate and. 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. The bicinchoninic acid bca protein assay primarily relies on two reactions. The kit is an optimized mix and read assay format that is compatible with hts liquid handling instruments.
Thermo scientific pierce protein assay technical handbook. Us4839295a measurement of protein using bicinchoninic. Although the mechanism of color formation for the lowry assay is similar to that of. Quantumprotein bicinchoninic acid protein assay1,2,3 kit is formulated for the colorimetric detection and quantitation of total protein. Development and validation of a hplc analytical assay. Bicinchoninic acid bca method an example of an indirect. 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. The detergentcompatible method provides for accurate determination of protein. Qubit, bradford or bicinchoninic acid in related conceptions. 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.
When it is necessary to determine the total protein concentration in a. Unfortunately this type of reductometric assay is not speci. The purple colored cubicinchoninic acid2 3 complex displays a strong absorbance at 562 nm which is proportional to protein concentration over a broad. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. Protein determination is one of the most common operations performed in biochemical research.
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. 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 bca is stable, it is incorporated in the reagent formulation at. 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. Insitu microvolume bicinchoninic acid protein assay. 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 remains one of the most popular protein assay methods worldwide. Chemistry of protein assays thermo fisher scientific us. This microbiological method is used to quantitatively measure serum and red blood cell rbc folate. Bicinchoninic acid bca assay in low volume sciencedirect. Tools and reagents for improved quantitation of total or. 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. This procedure is very applicable to microtiter plate methods. An adaptation of pierces bca protein assay reagent kit product no. The bca protein assay is used for quantitation of total protein in a sample. Adaptation of the bicinchoninic acid protein assay for use with microtiter plates and sucrose gradient fractions. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from. The color produced is stable and increases in a linear fashion over a broad working range of increasing protein concentration. The bicinchoninic acid bca 2 assay is a colorimetric method that is commonly employed to estimate the concentration of protein in a sample. 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.
The bicinchoninic acid method is faster and easier than lowry, with. The pierce micro bca protein assay reagent kit is a detergentcompatible bicinchoninic acid formulation for the colorimetric detection and quantitation of total protein. 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. Bicinchoninic acid bca protein assay, gbiosciences vwr. Competitive binding to cuprous ions of protein and bca in. Comparisons to a minibca assay workflow indicate that insitu analysis is more accurate. Using the micro bca protein assay kit pierce company. Since then it has become the most popular method for colorimetric detection and quantitation of total protein.
Piercea protein assay kit bc thermo fisher scientific. The bca assay is, however, more sensitive to interference from reducing sugars, possibly avidin gelatin e c 0 n n w u z q m o h m q bsa gelatin 200 400 800 1200 protein concentration kgml protein assay using bicinchoninic acid because the protocol allows the sugars more time to reduce cu to cu. This definition appears frequently and is found in the following acronym finder categories. Improvements in sensitivity are evident relative to standard protocols using 20. The two assays are of similar sensitivity, but since bca is stable under alkali conditions. Single amino acids and dipeptides do not give the biuret reaction, but. View product specific information, msds, references and buying faq. Decreases camp levels in native cultured cerebellar granule cells. The method is relatively easy to perform, reliable, and. The bicinchoninic acid bca assay, first described by smith et al. 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. 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.
Modified bicinchoninic acid assay for accurate determination of. Providers we are changing the way providers access and engage customers to streamline the sale and. Insitu microvolume bicinchoninic acid protein assay peter brescia, msc. Tools and reagents for improved quantitation of total or specific proteins. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. Best bicinchoninic acid bca protein quantification. We determined the protein concentration by the bicinchoninic acid assay sigma, using bovine serum albumin as. The bca is used for the same reasons the lowry is used.
The bicinchoninic acid bca assay is available in kit form from pierce rockford, ill. 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 bca is one of the most popular reagents for colorimetric protein determinations. 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 bca protein assay was introduced by smith, et al. The a 600nm response was found to be linear at least over the range 0. 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. This protocol describes a method of quantifying protein that is a variation of the lowry assay. Bicinchoninic acid assay an overview sciencedirect topics. A stock bca solution contains the following ingredients in a highly alkaline solution with a ph 11.
The absorbance at 562 nm of the cuibicinchoninate2 complex is directly proportional to protein concentration. Bicinchoninic acid assay science exchange lets you compare quotes from over 10 leading service providers. 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. Callan from the lankenau hospital research institute, philadelphia received for publication, june 17, 1938 the best method at present available for the exact titrimetric. Bca is less variable and less susceptible to detergents than bradford, while bradford assay has a simpler and faster procedure. The assay was run automated as the absorbance change at 100 seconds and 400 seconds. The assay deviates from linearity at analyte concentrations beyond this range. Pierce bca protein assay kit thermo fisher scientific.
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