The bicinchoninic acid assay, also known as the smith assay, after its inventor, paul k. 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. 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. 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, similar to lowry protein assay, bradford protein assay or biuret reagent. The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex. Bca bicinchoninic acid protein assay is watersoluble and strongly absorbs light at 562nm in a linear fashion with increasing protein concentration. Copper solution, for example, for 10ml working solution combine 5ml assay. Mucf protein was quantitated by bicinchoninic acid assay pierce chemical co. An adaptation of pierces bca protein assay reagent kit product no. Total protein content was measured by a standard bicinchoninic acid assay bca assay. In particular, it is insensitive to detergents such as triton x100 and sds 1 %. 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. Bca bicinchoninic acid protein assay kit with bsa standards.
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. The bca protein assay is used for quantitation of total protein in a sample. Quantitation of total protein content is a measurement common to many applications in basic science and clinical research. Microplate procedure linear working range of 240 gml. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed. Standard protein quantification methods, such as bicinchoninic acid assay bca will suffice. Following bicinchoninic acid assay bca quantification, 10. The principle of lowry assay, biuret assay, and bicinconinich bca assay protein assays duration. Bicinchoninic acid bca protein assay smith rice university.
The purplecolored reaction product of this assay is formed by the chelation of two molecules of bca with one cuprous ion. Insitu microvolume bicinchoninic acid protein assay february 26. These keywords were added by machine and not by the authors. Competitive binding to cuprous ions of protein and bca in the. The bicinchoninic acid bca assay, first described by smith et al. Compared to native protein absorbance at 280 nm, bca has improved sensitivity and good specificity for protein with minimal interference from. Ammonium sulfate sodium azide standard assay bicinchoninic acid sodium tartrate. Best bicinchoninic acid bca protein quantification. Quantumprotein bicinchoninic acid protein assay1,2,3 kit is formulated for the colorimetric detection and quantitation of total protein. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent.
The two assays are of similar sensitivity, but since bca is stable under alkali conditions. Bicinchoninic acid assay an overview sciencedirect topics. Unfortunately this type of reductometric assay is not speci. 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. Box 117 industriezone iii rockford, il 61105 industrielaan 27 usa 9320 erembodegemaalst 815. Bicinchoninic acid kit and bovine serum albumin bsa were obtained from sigmaaldrich. Copperii sulfate pentahydrate 4% solution, catalog number c2284 reagent b is a 25 ml solution containing 4% wv copperii sulfate pentahydrate. The size distribution and concentration of purified omvs were determined by. Using the micro bca protein assay kit pierce company, we measured the. Bicinchoninic acid assay science exchange lets you compare quotes from over 10 leading service providers. Measurement of protein using bicinchoninic acid sciencedirect. This protocol describes a method of quantifying protein that is a variation of the lowry assay. Sodium azide sodium acetate ammonium sulfate standard assay bicinchoninic acid.
This procedure is very applicable to microtiter plate methods. Pierce bca protein assay kit, sufficient reagents for 500 testtube or 5000 microplate assays. 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. Direct nonisotopic assay of 3methylglutaconylcoa hydratase in. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. Pdf competitive binding to cuprous ions of protein and bca in. The basis for this assay is the reduction of copper in alkali by protein biuret reaction in the presence of the chromogenic agent bicinchoninic acid. Bicinchoninic acid protein assay kit is a formulation based on bicinchoninic acid for the rapid and sensitive detection and quantitation of total protein content. First is the biuret reaction, whose faint blue color results from the reduction of cupric ion to cuprous ion. Expired lifetime application number us07053,708 inventor paul k. More sensitive than lowry and less subject to interferences.
Bca protein assay reagent bicinchoninic acid from thermo. This reagent forms the basis of an analytical method capable of monitoring cuprous ion produced in the. Protein quantitation based on bicinchoninic acid bca is simple, sensitive, and tolerant to many detergents and substances known to interfere with the lowry method. The bicinchoninic acid bca assay is available in kit form from pierce rockford, ill. Suitability this product is commonly used for protein determination by the bicinchoninic acid bca method. Bicinchoninic acid protein assay kit bca1 technical. Dilution scheme for standard test tube protocol and microplate. When ready to run the assay, equal amounts of protein should be loaded per condition. 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. Bicinchoninic acid bca method an example of an indirect multi. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from nonionic detergents and buffer salts. To evaluate the performance of each of the purification methods performed earlier, purified omvs were evaluated by several methods as summarized in table 1. 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. 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.
Bca protein assay reagent bicinchoninic acid youtube. Insitu microvolume bicinchoninic acid protein assay. 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 protein assay, gbiosciences vwr. Linear working range for bsa is 20 2000 gml for the bicinchoninic acid protein assay. Nov 03, 2010 since the original work of smith et al. The bicinchoninic acid method is faster and easier than lowry, with. Suitability this product is commonly used for protein determination by the. The bicinchoninic acid bca protein assay primarily relies on two reactions. A new protein assay may be adequate in different experiments beyond or instead of the wellknown standard protocols e. Using the micro bca protein assay kit pierce company, we measured the absorbance at 562 nm of bsa solutions with different concentrations of protein. Us4839295a measurement of protein using bicinchoninic acid. The first format was the minibca assay recommended by the nanodrop technical manual 2.
The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. Packaging 1 l in poly bottle also available as part of the bicinchoninic acid kit for protein determination. Biomedical and biological sciences 61,877 views 24. Pdf measurement of protein using bicinchoninic acid m. Combine 50 parts bca solution with 1 part copper solution, for example, for 10ml working. Synonyms bicinchoninic acid protein assay reagent a intended use refer to the instruction booklet for proper and intended use. An adaptation of pierces bca protein assay reagent kit product. Total dna content was determined by measuring the absorbance at 260 nm.
Micro bca reagent b mb, 240 ml, containing bicinchoninic acid 4. Pierce bca protein assay kit thermo fisher scientific. 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. The bicinchoninic acid bca assay for protein quantitation. We determined the protein concentration by the bicinchoninic acid assay sigma, using bovine serum albumin as the calibrator. 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. Piercea protein assay kit bc thermo fisher scientific.
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 bca assay was demonstrated using two microvolume formats. Bicinchoninic acid bca protein assay photometric method reading at 562 nm. Micro bicinchoninic acid bca protein assay gbiosciences. Add 10l of bsa standards stored in 80c freezer to each of the. Qubit, bradford or bicinchoninic acid in related conceptions. The pierce micro bca protein assay reagent kit is a detergentcompatible bicinchoninic acid formulation for the colorimetric detection and quantitation of total protein. The bicinchoninic acid method is faster and easier than lowry, with much greater tolerance to interference from. Herein, we used the pierce bca protein assay kit thermo scientific, catalog no. When it is necessary to determine the total protein concentration in a. The thermo scientific pierce bca protein assay kit is a. Bicinchoninic acid protein assay kit bca1 technical bulletin. Pdf although bicinchoninic acid bca has been widely used to determine. Otherwise, contact supplier for specific applications.
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