Reagents in the bca thermo scientific a javascript. Standard 2 ml assay protocol this is the standard assay that can be performed in a test tube. Add 10l of bsa standards stored in 80c freezer to each of the appropriate wells. Equilibrate all reagents and samples to room temperature, then mix each thoroughly but gently to avoid micro bubbles. Incompatible substances amount compatible buffer systems nacetylglucosamine 10 mm in pbs, ph 7. The primary pair was used for generatingdiluent the data shown in the assay performance section and is included in the rplex human bca 1blc assay. The micro bicinchoninic acid bca protei n assay primarily relies on two reactions. Pdf is a hugely popular format for documents simply because it is independent of the hardware or application used to create that file.
Studies of the bca assay for determination of total protein. Add 1ml working solution to each tube, seal and vortex to mix. 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. Bca bicinchoninic acid protein assay fanglian he department of. The assay is linear over the widest range of protein concentration between 252000 gml. 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. 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. The absorbance value is proportional to the protein concentration. This is part of the free pocketlab mobile application. Bicinchoninic acid bca assay or smith assay is a copperbased colorimetric assay for total protein quantification.
The following sections provide examples of assay files for common types of assays identified across nanotechnology resources. Immunofluorescence microscope to analyze the subcellular localization of the mutant agt is inconclusive. Do triplicates of all samples including the standards. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this assay has the advantage that it can be carried out as a onestep process compared to the two steps needed in the lowry assay.
Home contact us covid19 is an emerging, rapidly evolving situation. Plate map with a standard curve from 0 to 100 gml with samples and standards in duplicate. Therefore, the protein concentration can be calculated according to the od value. The standard protocol permits either a 30minute incubation at 37c or a 2hour incubation at room temperature. The standard protocol can be performed in three different formats, 5 ml and a 1 ml cuvette assay, and a 250 l microplate assay. The bicinchoninic acid bca assay, first described by smith et al. The watersoluble bca copper complex absorbs more strongly than the peptidecopper complex, increasing the sensitivity of the biuret assay from 5 mgml to 0. Methods of determining protein concentrations in a sample are disclosed comprising the steps of combining the sample with a bca assay reagent in a sample container, placing the sample container into a.
The linear range of these assays for bsa is 1251,000 gml, whereas with gammaglobulin the linear range is 1251,500 gml. The purplecolored reaction product of this assay is formed by the chelation of two molecules of. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this assay has the advantage that it can be. An adaptation of the traditional bca assay, the rapid gold bca protein assay kit has been optimized to develop in 5 minutes at room temperature.
Download bca protein assay protocol thermo scientific pdf. A modification of the standard bca protocol is provided that allows sensitive and reliable protein determinations in a matter of seconds utilizing a microwave oven to irradiate the samples. Bca protein assay protocol a solutions and reagents kit contents. The bsa protein standard consists of a known concentration of bovine serum albumin, and the unknown sample is the solution to be assayed. The pathlength in each assay is dependent on the assay container cuvettes or multiwell plates andor the reaction volume.
Abstract the bca protein assay is used for quantitation of total protein in a sample. Protein concentrations between 5250 gml can be determined with this protocol. Pdf the bca assay is a colorimetric method for estimating protein concentration. Piercea protein assay kit bc thermo fisher scientific. By michelle rae uy 24 january 2020 knowing how to combine pdf files isnt reserved. The remaining kit components should be stored at room temperature. The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex. Luckily, there are lots of free and paid tools that can compress a pdf file in just a few easy steps. Frederick national laboratory for cancer research standard. Pierce bca protein assay kit, sufficient reagents for 500 testtube or 5000 microplate. Bca assay can be performed in a microtiter plate format or test tube format. Bca assay high levels of ionic detergents require the addition of the ionic detergent compatibility reagent idcr. An oversized pdf file can be hard to send through email and may not upload onto certain file managers.
In 96well plates, the relationship between protein content and absorbance is nearly linear over a wide range. 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. Read on to find out just how to combine multiple pdf files on macos and windows 10. Each method has its advantages and disadvantages table 1. In the bca scientific protocol place by leaving comments, feedback or suggestions. The bca protein assay working range standard protocol. When stored properly, the kit is stable for 1 year. Protocol for bca protein assay using a nanodrop 8000. Bicinchoninic acid protein assay kit bca1 technical. A protein assay, such the bca protein assay, is an excellent tool for estimating the protein concentration of a sample.
The assay file will vary depending on the type of assay performed and protocol and technology type leveraged. The bca assay is a colorimetric method for estimating protein concentration. Pdf bicinchoninic acid bca assay in low volume researchgate. Bicinchoninic acid protein assay kit bca1 technical bulletin. Add 25 l of each bsa standard and protein samples into microtiter plate wells. Making a pdf file of a logo is surprisingly easy and is essential for most web designers. It is recommended that a list file be generated prior to starting the assay if many samples are to be measured. Prepare enough fresh working reagent for all standards. The bca protein assay is used to determine the protein concentration of an unknown sample.
Download bca protein assay protocol thermo scientific doc. The bicinchoninic acid bca assay for protein quantitation. This means it can be viewed across multiple devices, regardless of the underlying operating system. Adobe systems is a software manufacturer that has created many document and multimedia editing programs. Once youve done it, youll be able to easily send the logos you create to clients, make them available for download, or attach them to emails in a fo. 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. Tools and reagents for improved quantitation of total or. No one reagent can be considered to be the ideal or best protein assay method. Pipette 50l of each standard and protein samples into an appropriately labeled tube. The bca protein assay kit contains all the reagents required to carry out over assays in a 96 well format. Module and the bca protein assay is 5 gml figure 1 and the maximum is 2 mgml figure 2. Thermo scientific pierce micro bca protein assay kit standard protocol. The 96 well plate assay is for those who wish to perform the bradford assay in plate format. Studies of the bca assay for determination of total.
Bicinchoninic acid assay an overview sciencedirect topics. In addition, the kit also contains a detailed protocol and references for use. The test tube assay requires a larger volume of protein sample 50 l. Biovisions reducing agent compatible bca protein assay kit is the only protein estimation kit on the market that is compatible with. Linear regression curve fit is used to determine unknown concentrations. The assay can be completed in 5 minutes at room temperature eliminating the need to wait or expose samples to elevated temperatures. The enhanced protocol requires a 60c incubation for 30 minutes. Bca working reagent reagent a mixed with reagent b is stable for one day. Searching for a specific type of document on the internet is sometimes like looking for a needle in a haystack. A pdf portable document format is a widely popular type of document format created by adobe. Micro bca protein concentration assay use the thermo scientific micro bca protein assay kit. 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. These and others changes like the bca working reagent to protein sample ratio affect the net absorbance values. The biorad protein assay is a dyebinding assay in which a differential color change of a dye occurs in response to various concentrations of protein.
Safety data sheet bicinchoninic acid bca protein assay. The bca assay is suitable for comparative assays with diverse potential polysaccharide substrates and can be used either for specific activity e. If the total protein concentration in the samples is high 2,000. Pdf file or convert a pdf file to docx, jpg, or other file format. The bca protein assay from gbiosciences comprises two assay formats based upon the dynamic range required to measure the concentration of the protein of interest. The rapid gold bca protein assay produces an orangegold colored reaction product, which is formed by the chelation of 2 molecules of the chelator with 1 cuprous ion. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions.
The thermo scientific micro bca protein assay kit is a detergentcompatible bicinchoninic acid formulation for the colorimetric detection and quantitation of. Thermo scientific pierce protein assay technical handbook. These instructions are a step by step procedure on how to perform the standard assay. A purple colored reaction product is formed by the chelation of two molecules of bca with one cuprous ion. Bca protein assay kit reducing agent compatible rev. The microassay is for samples with low protein concentrations. Preparation of standards and working reagent required for both assay.
Procedure in the standard assay, 20 parts of the bca working reagent are then mixed with 1 part of a protein sample. Bca enhanced protocol assay was performed on the modulus using the. The bca protein assay is a popular method for colorimetric detection and quantitation of total protein. 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. If another format is required, extrapolate the volume of samples and reagents.
The determination of protein concentration is frequently required in biochemical work. However, the effect of interfering substances is minimized because the ratio of bca working reagent to protein sample is 20. Dilute the contents of one albumin standard bsa ampule into several clean vials, preferably using the same diluent as the samples. A pdf file is a portable document format file, developed by adobe systems. The rplex human bca 1blc antibody set has been tested with the following diluent pairs. Pierce bca protein assay kit thermo fisher scientific. Alternate diluent pairs may be considered for use with rplex antibody sets, based on the sample type.
It can be performed either in a 1ml cuvette assay or in microwells. To combine pdf files into a single pdf document is easier than it looks. Bradford protein assay protocol university of san diego. Assay cascade protocols nanotechnology characterization lab ncl. The bca protein assay reducing agent compatible standard protocol. Bicinchonininsaure bca einen violetten komplex bilden. This procedure uses 1 ml of a protein sample and 1 ml of the mixed quantipro w orking reagent.
A purple colored reaction product is formed by the chelation of two molecules of bca. The standard protocol permits either a 30minute incubation at 37c or a 2 hourincubation at room temperature. The reason for a pdf file not to open on a computer can either be a problem with the pdf file itself, an issue with password protection or noncompliance w the reason for a pdf file not to open on a computer can either be a problem with the. The enhanced protocol requires a 30minute incubate at 60c. The choice among available protein assays is usually based on the compatibility of the protein assay method with the samples. For the enhanced protocol, incubate at 60c for 60 minutes. Methods of determining protein concentrations in a sample are disclosed comprising the steps of combining the sample with a bca assay reagent in a sample container, placing the sample container into a microwave oven, irradiating the sample and measuring an absorbance value at 562 nm for the sample. By modifying the protocol you can change the range to 202000 gml. For the 96 well plate assay, 8 parts of the bca working reagent are mixed with 1 part of a protein sample.
Before reading the bca thermo scientific of samples is essential for everyone. The bca protein concentration kit is an ideal protein quantification method which is superior to the lowry method. This article explains what pdfs are, how to open one, all the different ways. Assay protocols the following procedures describe two assay protocols. Our improved formulation, the bc assay, provides significant benefits over existing colorimetric assays, namely sensitivity, reagent stability, and detergent compatibility.
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