The takara bca protein assay kit is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. The bca protein assay was performed following the manufacturers protocol. Studies of the bca assay for determination of total. Being present a electricity, proteins migerate towards the negative. Incompatible substances amount compatible buffer systems nacetylglucosamine 10 mm in pbs, ph 7. This unit describes how to perform commonly used protein assays, e. 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. Pdf bicinchoninic acid bca assay in low volume researchgate. According to the manufacturers protocol this protein assay is linear in the range of 0. This site is like a library, you could find million book here.
The bicinchoninic acid bca protein assay primarily relies on two reactions. I want to do a bca protein microplate assay pierce but our plate reader only has a 595 nm filter. The standard protocol permits either a 30minute incubation at 37c or a 2 hourincubation at room temperature. Preparation of standards and working reagent required for both assay procedures. The reaction reaches 90% of its maximum color development within 15 minutes thereby. Smith at the pierce chemical company in 1985 smith et al.
Biotek sample files, 08jun11, bca protein assay with protein concentration curve standard curve and calculation of unknown concentrations. Download a modulus method for bca protein assay book pdf free download link or read online here in pdf. The manufacturer of the bca kit advises to use a 562 nm filter, or at most a 590 nm filter. Thermo scientific pierce protein assay technical handbook. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay that is. The thermo scientific pierce bca protein assay kit is a detergentcompatible dilution scheme for standard test tube protocol and microplate procedure proteins.
Be aware of any hazardous compounds, take precautions where necessary and dispose of any waste in the appropriate manner. For more information about sbi products and to download manuals in pdf format. The bradford assay was successfully performed on the bmg labtech microplate reader. A typical bca assay standard curve with samples ranging from 0. Lowry protein assay an overview sciencedirect topics. 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. The bca assay is more sensitive and applicable than either biuret or lowry procedures. Assays for determination of protein concentration olson.
If the total protein concentration in the samples is high 2,000. Read online a modulus method for bca protein assay book pdf free download link book now. The bicinchoninic acid bca 2 assay is a colorimetric method that is commonly employed to estimate the concentration of protein in a sample. Download the pdf or microsoft word versions of this protocol. 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. Compatibility of pierce bca protein assay with promega lysis. Bca versus biorad protein assay what are the advantages of bca that all the world uses it.
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 assay can be completed in 5 minutes at room temperature eliminating the need to wait or expose samples to elevated temperatures. The bca protein assay reducing agent compatible standard protocol. Download our protein assays and bioassays handbooks. Bca assay high levels of ionic detergents require the addition of the ionic detergent compatibility reagent idcr.
Quick start bradford protein assay harvard university. It is recommended that the bca compatibility reagent be used to decrease interference from reducing agents, chelators, detergents, and other common ingredients found in most lysis buffers. The bicinchoninic acid bca assay, first described by smith et al. The amount of purplecolored product formed during the reaction can be correlated to the amount of protein in the bca protein assay. Bicinchoninic acid bca assay in low volume sciencedirect. Add 25 l of each bsa standard and protein samples into microtiter plate wells. 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. Sulforhodamine b colorimetric assay for cytotoxicity screening. These and others changes like the bca working reagent to protein sample ratio affect the net absorbance values. This protocol works best from a frozen or fresh exosome pellet or in a highly. The sulforhodamine b srb assay is used for cell density determination, based on the measurement of cellular protein content.
This protocol works best from a frozen or fresh exosome pellet or in a highly concentrated solution about 107 exosomesl, corresponds to a protein concentration of 2gl. Protein assay compatibility table from our website. The standards used for the bca assay usually bovine serum albumin should contain the same amount of sds as the samples. In test tubes or 96well plates, the relationship between protein concentration and absorbance is nearly linear over a.
The standard protocol permits either a 30minute incubation at 37c or a 2hour incubation at room temperature. Add approximately 20100 g exosomal protein per reaction. A modulus method for bca protein assay pdf book manual. This protocol describes a method of quantifying protein that is a variation of the lowry assay. Bicinchoninic acid bca protein assay, gbiosciences vwr. The reaction is similar to the welldocumented lowry1 assay, but with the following improvements.
The two assays are of similar sensitivity, but since bca is stable under alkali conditions. The biorad dc protein assay is a colorimetric assay for protein concentration following detergent solubilization. Certain substances interfere with the pierce bca assay kit. The thermo scientific pierce bca protein assay kit is a detergent compatible formulation based on bicinchoninic acid bca for the. Bca assay can be performed in a microtiter plate format or test tube format. The enhanced protocol requires a 60c incubation for 30 minutes. The enhanced protocol requires a 30minute incubate at 60c. Bicinchoninic acid protein assay kit bca1 technical bulletin. The absorbance at 562nm was read in a spectramax 250 plate reader. A 25l sample volume is recommended for the microplate procedure of the bca protein assay. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. The principle of this technique is quite similar to the lowry protein assay, which is one of the alternative protein quantification methods. 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.
Streamline bcabased protein quantitation molecular devices. Pierce bca protein assay kit, sufficient reagents for 500 testtube or 5000. The pathlength in each assay is dependent on the assay container cuvettes or multiwell plates andor the reaction volume. The bicinchoninic acid bca assay method for the determination of protein has been investigated for its utility in measuring. Mar 27, 2020 download a modulus method for bca protein assay book pdf free download link or read online here in pdf. Each assay was performed in duplicate, and the results are presented as the average of duplicates. Piercea protein assay kit bc thermo fisher scientific. You can do a protein assay with a small aliquot of the precipitated exosomes to determine how. The bicinchoninic acid bca protein assay is a highly sensitive colorimetric assay the thermo scientific pierce bca protein assay kit is a detergentcompatible dilution scheme for. Pierce bca protein assay kit thermo fisher scientific. The method described here has been optimized for the toxicity. An adaptaon of the bca protein assay kit, the micro bca kit has been opmized for use with dilute protein samples 0. The bca assay plate was read at 562 nm, the original wavelength setting in the protocol, while for the bca rapid gold assay the wavelength setting was changed to 480 nm prior to reading the plate.
The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. 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. Tools and reagents for improved quantitation of total or. The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex. 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 primary focus of the unit is assay selection, emphasizing sample and buffer compatibility.
The bca protein assay kit is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Sdspage, with full name of sodium dodecyl sulfate polyacrylamide gel electrophores, is the most widely used technique to separate proteins from complicated samples of mixture, plays key roles in molecular biology and wide range of subfield of biological research. All books are in clear copy here, and all files are secure so dont worry about it. Aug 17, 2006 the sulforhodamine b srb assay is used for cell density determination, based on the measurement of cellular protein content. Has anyone successfully used a 595 nm filter for this assay.
The bicinchoninic acid bca assay is a protein quantification technique that was first demonstrated by paul k. This was done by analyzing the precision, accuracy, linearity and range of the method. Please see the attached protocol for additional details. Bsa standard curves were generated by softmax pro software, and a quadratic curve fit was applied, as recommended in the assay kit user guides. 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. I have found the assay to be fairly sensitive and reliable, however a few points should be noted. Biochemical analysis of proteins relies on accurate quantitation of protein concentration. 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. If you treat them the same way as the samples, that is by adding the sds. The remaining kit components should be stored at room temperature. 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. The bca protein assay is used for quantitation of total protein in a sample.
Because of its homogeneous and fast nature, the assay is a preferred method to determine the protein concentration of samples. For more product information, or to download a product instruction booklet, visit. Bca enhanced protocol assay was performed on the modulus using the. The bicinchoninic acid bca protein assay is based on a twostep reaction in one tube. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this assay has the advantage that it can be. Studies of the bca assay for determination of total protein. 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 protein assay with protein concentration curve. Bca protein assay reagent kit, sufficient reagents for 250 test tube or 2,500 microplate assays. Compatibility of pierce bca protein assay with promega.
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