A colleague of mine recommended that i use the bca assay, because he had more success with this assay in the past. Cellular assays have always been a powerful tool in the research lab. Available in tube and microplate formats, these reducing agent compatible rac bca kits extend the already broad reagent compatibility of the bca protein assay protocol to include. Lowry protein assay an overview sciencedirect topics.
Additional information and online ordering used in more labs than any other detergentcompatible protein assay, pierce bca reagents provide accurate determination of protein concentration with most sample types. Lowry method is the most cited protein assay in the literature adaptable for use with microplates. The results for protein content from the bradford assay is low compared to a combustion method in the same biomass 35% from combustion, 20% from bradford assay. Thermo scientific pierce microplate bca protein assay kit. Both micro bca and pierce bca assays can quantify protein via either microplate or test tube procedure. Bca protein assay kit ab102536 provides a simple, rapid, detergent tolerant up to 5% procedure for determining the concentration of proteins in solution. A glomaxmulti jr method for bca protein assay introduction the glomaxmulti jr and the pierce bca protein assay provide a simple method for quantifying protein over a large range. Compared to most dyebinding methods, the bca assay is affected much less by protein compositional differences, providing greater concentration accuracy. They are incredibly versatile, and can be designed to measure virtually any cellular or biochemical function. 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. Protein mass spectrometry is a relatively new and developing method for protein quantitation. Disposable standard and semimicro plastic cuvettes ideal for these protein assays are also available. The bca protein assay is a copperbased protein assay and is also known as the smith assay because it was introduced by paul k. Prazises, mit detergens kompatibles assayreagenz grun bis blau, a 562 nm zur messung.
The kit has been optimized for determining total protein content in low protein concentration samples 0. The principle of lowry assay, biuret assay, and bicinconinich bca assay protein assays duration. How to calculate sample protein based on standard protein. Besides protein characterization, an important step in the proteomic analysis is the possibility to quantify a specific protein. Conducting the bicinchoninic acid bca assay directly after a coupling reaction using 1ethyl33dimethylaminopropyl carbodiimide edc and nhydroxysuccinimide nhs chemistry produces significant errors. Thermo scientific pierce bca protein assay kit fisher. Thermo scientific pierce bca protein assay kit remains one of the most popular protein quantitation methods worldwide. Pierce bca protein assay kit thermo fisher scientific. Furthermore, the volume of reagents can be reduced, and it can be performed in 96well plates. Jun 19, 2018 the principle of lowry assay, biuret assay, and bicinconinich bca assay protein assays duration. Thermo scientific pierce 660 nm protein assay overview highlights typical response curves 14 bcabased protein assays 15 chemistry of the bca protein assay 15 advantages of the bca protein assay 16 disadvantages of the bca protein assay 17 bca, reducing agent compatible 18. The intensity of the colored reaction product is a direct function of protein amount that can be determined by comparing its absorbance value to a standard curve.
Thermo scientific pierce bca protein assay kit bca. Feb 14, 2014 accurately measure the total protein concentration in your sample with this twocomponent, highprecision, detergentcompatible assay reagent set. The pierce bca assay can be used to assess yields in whole cell lysates and affinitycolumn fractions, as well as to monitor protein contamination in industrial applications. Thermo scientific pierce bca protein assay kit bca protein. Pierce bca protein assay kit university of arizona. Thermo scientific pierce bca proteinassay fisher scientific. 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 method has greater proteintoprotein variability than the bca assay. Compared to most dyebinding methods, the bca assay is affected much less by protein compositional differences, providing greater proteintoprotein uniformity. Biorad offers four colorimetric protein assay kits for the measurement of protein levels using modifications of the bradford and lowry assay method.
Many techniques have been introduced and implemented for protein quantitation by mass spectrometry. Pierce bca protein assay kits and reagents, thermo scientific. Thermo scientific pierce bca protein assay kit reagent a. To determine the total protein concentration of a sample a bsa standard curve has to be set. The bca protein assay is a popular method for colorimetric detection and quantitation of total protein. Thermo scientific pierce 660 nm protein assay overview highlights typical response curves 14 bca based protein assays 15 chemistry of the bca protein assay 15 advantages of the bca protein assay 16 disadvantages of the bca protein assay 17 bca, reducing agent compatible 18 bca protein assay 19 micro bca protein assay 20. The bicinchoninic acid bca protein assay is suitable for quantifying protein solutions in 1ml assays or in microwells. The pierce micro bca protein assay reagent kit is a detergentcompatible bicinchoninic acid formulation for the colorimetric detection and quantitation of total protein. The pierce bca protein assay can be used to assess yields in whole cell lysates and affinitycolumn fractions, as well as to monitor protein contamination in industrial applications. A brief description of the principle and the choice of the experiment is as follows bca principle.
The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Competitive binding to cuprous ions of protein and bca in. An adaptation of pierces bca protein assay reagent kit product no. Piercea protein assay kit bc thermo fisher scientific. The pierce protein assay is a method of protein quantification. However, because this method is laborious and typically yields. The pierce bca assay can be used to assess yields in whole cell lysates, affinitycolumn fractions, purified proteins samples, as well as to monitor protein contamination in industrial applications. Accurately measure the total protein concentration in your sample with this twocomponent, highprecision, detergentcompatible assay reagent set. Pierce bca reagents provide accurate determination of protein concentration with most sample types encountered in protein research. I have a technical question regarding the bca assay. Measuring protein concentration using the bca protein assay with. The bca bicinchoninic protein assay is a widely used method for colorimetric detection and quantitation of total protein in a solution. This special adaptation of the popular pierce bca protein assay method enables accurate protein concentration measurement for samples containing dtt, 2me or tcep. Pierce bca protein assay kit is a twocomponent, highprecision, detergentcompatible protein assay for determination of protein concentration.
The bca protein assay or smith assay is a biochemical assay for determining the total level of protein in a solution. Pierce offers seven colorimetric assays for detection and quantitation of total protein. Protein assay kits and cuvettes life science research. This procedure is very applicable to microtiter plate methods. The bicinchoninic acid bca assay is a protein quantification technique that was first demonstrated by paul k. The thermo scientific pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. This is the protocol to be used in the determination of total protein in brain tissue by the pierce bca method. Oct 22, 2007 the bca protein assay kit from thermo scientific pierce product no. It provides quick estimation of the protein amount in a given sample. Bca method has less proteinprotein variability than the bradford assay.
Lowry method is the most cited protein assay in the. Micro bca protein assay kit pierce micro bicinchoninic acid. This method may be preferred for samples containing detergents or other reducing agents. The bca protein assay is a simple and reliable protein quantification method. Pierce bca bicinchoninic acid reagents provide accurate determination of protein concentration with most sample types encountered in protein research. The amount of protein present in a solution can be quantified by measuring the absorption spectra and comparing with protein solutions with known concentrations. The bca protein assay working range standard protocol. This reducing agentcompatible rac version of the thermo scientific pierce bca protein assay provides accurate determination of protein concentration in samples that contain dithiothreitol dtt, 2mercaptoethanol bme, tcep and other disulfide reducing agents. Bca protein assay kit from thermo scientific pierce. Used in more labs than any other detergentcompatible protein assay, pierce bca reagents provide accurate determination of protein concentration with most sample types.
The bicinchoninic acid bca assay is available in kit form from pierce rockford, ill. The bca protein assay kit from thermo scientific pierce product no. Pierce bca protein assays have a unique advantage over. The kit is sensitive to copper reductant and chelator instead of to both ionic and nonionic detergents. The micro bca protein assay kit provides an ultrasensitive method to detect and quantify protein concentration. Difference between bca and bradford method to determine.
Thermo scientific pierce bca proteinassay protein assay kit. Bca assay openwetware bca protein assay kit test tube reducing agent. Smith at the pierce chemical company in 1985 smith et al. Micro bca protein assay 18 bca, reducing agent compatible see page 24. Micro bca protein assay kit pierce micro bicinchoninic. Correction in bicinchoninic acid bca absorbance assay to.
Here we present a correction for the quantification of gelatin in the supernatant sn following gelatin conjugation to polymer microparticles using edc and nhs chemistry. The thermo scientific pierce bca protein assay kit is a twocomponent, highprecision, detergentcompatible assay reagent set to measure a562nm total protein concentration compared to a protein standard. Reagent a, a carbonate buffer containing bca reagent, and reagent b, a cupric sulfate solution, which are combined to make an apple greencolored working solution that turns purple after 30 minutes at 37c in the presence of protein. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. The differences between the bca and bradford protein assay. Bca protein assay kit from pierce provides a simple method for quantifying protein over a large range. 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. Measuring the od at 595nm of the protein solution determines the final protein concentration. Bca protein assay reagent bicinchoninic acid youtube. Urine dip stick strips were purchased from euro diagnostic systems, india pvt. Compared to most dyebinding methods, the bca assay. Thermo scientific pierce bca proteinassay bca protein assay kit.
Bca protein assay for estimating detergent solubilized. The thermo scientific pierce 660nm protein assay is a quick, readytouse colorimetric method for total protein quantitation. Pierce bca protein assay kit, thermo scientific vwr. The glomax discover system and pierce bca protein assay provide a simple method for quantifying protein over a large range. The chemical treatment method, like that used in the bca protein assay reducing agent compatible, or the pierce 660 nm protein assay is generally preferred. The bca is used for the same reasons the lowry is used. Protein assay kits and cuvettes life science research biorad. The total protein concentration is determined by measuring the color change of the sample solution from green to purple which is proportional to the protein concentration. A stock bca solution contains the following ingredients in a highly alkaline solution with a ph 11. The two most commonly used methods for protein quantification are the bca method, bradford coomassie brilliant blue method. Pierce bca protein assays have a unique advantage over the coomassie dyebased assays bradfordthey are compatible with samples that contain up to 5% surfactants detergents, and are affected much less by protein compositional differences, providing greater proteintoprotein uniformity.
How to calculate sample protein based on standard protein curve in bca. The assay is reproducible, rapid and more linear compared to coomassiebased bradford assays and compatible with highconcentrations of most detergents, reducing agents and other commonly used reagents. Pierce bca protein assay kit, sufficient reagents for 500 testtube or 5000 microplate. Compared to most dyebinding methods, the bca assay is affected much less by protein compositional differences, providing greater protein to protein uniformity. The chemical treatment method, like that used in the bca protein assay reducing agent compatible, is generally preferred because, unlike protein precipitation, resolubilization of potentially hydrophobic proteins is not involved. Using the micro bca protein assay kit pierce company. Bsa bovine serum albumin is used as the standard and samples are. The standard pierce bca protein assay detects protein concentrations from 20 to 2,000. Bicinchoninic acid bca protein assay smith considerations for use. We use the pierce micro bca kit to determine protein concentrations in solutions that contain 0. This method has a fast detection speed and low proteintoprotein variability in comparison to the bca or coomassie bradford assays. Jul 23, 2014 the results for protein content from the bradford assay is low compared to a combustion method in the same biomass 35% from combustion, 20% from bradford assay. Protein determination by the bca method rice university. Thermo scientific pierce protein assay technical handbook.
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