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