For descriptions of symbols on product labels or product documents, go to. The assay is linear over the widest range of protein concentration between 252000 gml. 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. A purple colored reaction product is formed by the chelation of two molecules of. Pierce bca protein assay kit, sufficient reagents for 500 testtube or 5000. Pierce bca reagents provide accurate determination of protein concentration with most sample types encountered in protein research. Thermo scientific pierce protein assay technical handbook. All books are in clear copy here, and all files are secure so dont worry about it. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. For descriptions of symbols on product labels or product documents.
If precise quantitation of an unknown protein is required, it is advisable to select a protein. The thermo scientific pierce bca protein assay kit is a. In any protein assay, the best protein to use as a standard is a purified preparation of the protein being assayed. 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. The prediluted standards are conveniently packaged in. Bca is sensitive and has a broad dynamic range capable of measuring protein concentrations of 0. Add 25 l of each bsa standard and protein samples into microtiter plate wells. Jun 19, 2018 the principle of lowry assay, biuret assay, and bicinconinich bca assay protein assays duration. 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.
Bicinchoninic acid protein assay kit bca1 technical. Dilution scheme for standard test tube protocol and microplate procedure. Lower protein samples can be more easily quantified using higher temperatures and longer incubation times. The following are instructions for the microplate assay format of the pierce. Biovisions bca protein assay kit provides a colorimetric detection and. To determine the total protein concentration of a sample a bsa standard curve has to be set. Studies of the bca assay for determination of total protein in allergen extracts title swedish abstract the aim of this thesis has been to evaluate and to optimize the bca protein assay for quantification of total protein. The enhanced protocol requires a 60c incubation for 30 minutes. Studies of the bca assay for determination of total protein. This protocol makes use of thermo scientifics bca protein assay in a 96well tissue culture plate format to measure the amount of protein in small volumes of sample. Unfortunately we cannot switch to another protein assay as we. Compatibility chart for bca kit the amount listed is the maximum amount of material allowed in the protein sample without causing a noticeable interference, when 20 ul protein sample is used for bca assay as described in the standard assay. Biotek sample files, 08jun11, bca protein assay with protein concentration curve standard curve and calculation of unknown concentrations.
Four major protein assay protocols have been extensively used in the literature. The data sheet of the bca assay says that up to 5% of tween80 do not interfere with the assay, but we have problems even with 0. The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Bca total protein assay kit gene and cell technologies. Module and the bca protein assay is 5 gml figure 1 and the maximum is 2 mgml figure 2. Add 10l of bsa standards stored in 80c freezer to each of the appropriate wells. The bichinchoninic acid bca assay is used to estimate the total protein concentration in a sample. This is the protocol to be used in the determination of total protein in brain tissue by the pierce bca method. Pierce bca protein assay kit is a twocomponent, highprecision, detergentcompatible protein assay for determination of protein concentration. Bca protein assay with protein concentration curve standard. Pierce bca protein assay kit, sufficient reagents for 250 testtube or 2500 microplate assays. The quick start bradford protein assay is a simple and accurate procedure for determining the concentration of protein in solution. The quantipro bca assay kit is based on the same principals as the bca1 kit, except the quantipro bca kit gives a linear response from 0. Compare all available bca protein assays features of.
The bca protein assay kit can be used to measure the protein concentration of lysates or homogenates, in microplate format, prepared with the following buffers. Linear regression curve fit is used to determine unknown concentrations. Compared to most dyebinding methods, the bca assay is affected much less by protein compositional differences, providing greater proteintoprotein uniformity. A further advantage of the bca assay is that it is generally more tolerant to the presence of compounds that interfere. First documented in 1951, lowry assay was an oftencited general use protein assay for some time, but has been largely replaced by more robust assay protocols. Micro bca reagent b mb, 240 ml, containing bicinchoninic acid 4.
Bca protein assay kit, sufficient reagents for 250 test tube or 2,500 microplate assays. In comparison to the classical methods by bradford and lowry, the bca assay suffers less variability due to protein sequence, and is less prone to interference by nonprotein substances. The result is an extremely sensitive colorimetric protein assay in a test tube or microplate assay format. The glomax discover system and pierce bca protein assay provide a simple. Measuring protein concentration using the bca protein assay with.
Protein solutions are normally assayed in duplicate or triplicate. Bca assay can be performed in a microtiter plate format or test tube format. The standards used for the bca assay usually bovine serum albumin should contain the same amount of sds as the samples. This site is like a library, you could find million book here. Download a modulus method for bca protein assay book pdf free download link or read online here in pdf. Measuring the protein concentration of samples is essential for many protein assays, such as western blotting or zymography. Description tpro bca protein assay kit is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Bsa bovine serum albumin is used as the standard and samples are. Incompatible substances amount compatible buffer systems nacetylglucosamine 10 mm in pbs, ph 7. If you treat them the same way as the samples, that is by adding the sds. Bca protein assay for more information before performing the assay.
Lowry assay, biuret assay, bradford assay, and bca assay. Bicinchoninic acid protein assay kit bca1 technical bulletin. This product is for laboratory research only and not for diagnostic use. Coomassie bradford assay assay protocol utilizes small sample volumes available in both standard and microplate formats lowest proteintoprotein variability of any standard coomassie dyebased bradford assay faster than the bca or lowry protein assays ready to use. Scope all procedures are applicable to the bcgsc library core and library technology development groups iii. The best relative standard to use is one which gives a color yield similar to that of the protein. Biomedical and biological sciences 61,514 views 24. It is necessary to create a standard curve during each assay, regardless of the format used.
This unit also details highthroughput adaptations of the commonly used protein assays, and also contains a protocol for bca assay of total protein in sdspage sample buffer that is used for equal loading of sdspage gels, which is reliable, inexpensive, and. Read online a modulus method for bca protein assay book pdf free download link book now. Compared to most dyebinding methods, the bca assay is affected much less by protein compositional differences, providing greater protein to protein uniformity. Quantitative colorimetric peptide assay kit see related thermo scientific products. Piercea protein assay kit bc thermo fisher scientific. The microplate procedure affords the sample handling ease of a microplate.
This product is for laboratory research only and not for. Prepare three to five dilutions of a protein standard, which is representative of the protein solution to be tested. Bca bicinchoninic acid protein assay 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. Add 10l of unknown sample to each of the appropriate wells. Mar 27, 2020 download a modulus method for bca protein assay book pdf free download link or read online here in pdf. This colorimetric assay provides the high sensitivity and linearity associated with the bca assay, but in a fraction of the time it takes to perform a standard bca assay. Compare all available bca protein assays features of the bca protein assay kitreducing. The bicinchoninic acid bca assay, first described by smith et al. Measurement of protein using bicinchoninic acid sciencedirect.
The principle of lowry assay, biuret assay, and bicinconinich bca assay protein assays duration. The kit enables you to measure protein concentration in samples that contain the disulfide reducing agents. The two assays are of similar sensitivity, but since bca is stable under alkali conditions. The takara bca protein assay kit is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions.
The bicinchoninic acid bca assay for protein quantitation. Reagents bicinchoninic acid solution, catalog number b9643 reagent a is a 1,000 ml. The standard protocol permits either a 30minute incubation at 37c or a 2 hourincubation at room temperature. Of these, the test tube procedure requires a larger volume 0. It provides readytouse convenience by supplying the dye reagent at 1x concentration and two protein assay standards at seven prediluted concentrations. The bca protein assay kit can be used to measure the protein concentration of lysates or homog enates, in microplate format. The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex. In the absence of such an absolute reference protein, one must select another protein as a relative standard. Both the incubation time and temperature can be varied over a rather wide range. Quick start bradford protein assay harvard university. A purple colored reaction product is formed by the chelation of two molecules of bca with one cuprous ion.
The minimum concentration of bovine serum albumin bsa read by the glomaxmulti jr absorbance module and the bca protein assay is 5. Bca protein assay reducing agent compatible standard tube 1252,000 gml bca protein assay standard tube or microplate 202,000 gml enhanced tube 5250 gml micro bcaprotein assay standard tube 0. Protein assay standard curves and data processing fundamentals are discussed in detail. The bca assay has a linear concentration range between 2001,000 g of protein per milliliter. Bca enhanced protocol assay was performed on the modulus using the. This bca protein assay kit is a reducing agentcompatible version of our popular pierce bca protein assay. Pierce bca protein assay kit reducing agent compatible. The micro bca protein assay kit uses concentrated reagents and a protocol that utilizes an extended incubation time at an elevated temperature 60c, test tube procedure only.
The assay can be completed in 5 minutes at room temperature eliminating the need to wait or expose samples to elevated temperatures. Plate map with a standard curve from 0 to 100 gml with samples and standards in duplicate. The linear range of this microtiter plate assay is 0. Measuring the od at 595nm of the protein solution determines the final protein concentration. Bca assay can be performed in a microtiter plate format or test tube. 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. Pierce bca protein assay kit thermo fisher scientific. This was done by analyzing the precision, accuracy, linearity and range of the method. Microplate procedure linear working range of 240 gml. A modulus method for bca protein assay pdf book manual.
The enhanced protocol requires a 30minute incubate at 60c. 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. 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. Kit provides a convenient procedure for quantifying protein, and the assay protocol.
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