The pierce protein assay is a method of protein quantification. Pierce bca reagents provide accurate determination of protein concentration with most sample types encountered in protein research. Safety data sheet chandlergilbert community college. Protein assay versus the thermo scientific pierce 660 nm protein assay. 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. This unit describes how to perform commonly used protein assays, e.
Instructions pierce gst protein interaction pulldown kit. Urine dip stick strips were purchased from euro diagnostic systems, india pvt. The protein sample, irrespective of its composition, is measured through absorbance spectroscopy at 540 nm in conjunction with a k. Dc protein assay instruction manual for technical service call your local biorad office or in the u. Pierce assay has a significantly greater linear dynamic range than the bradford assay. The dye binding protein assay s are based on the binding of protein molecules to coomassie dye under acidic conditions. No action shall be taken involving any personal risk or without suitable training. They are all wellcharacterized, robust assays that provide consistent, reliable results. Protein concentration quantitation is an integral part of any laboratory workflow involving protein extraction, purification, labeling or analysis. The biuret assay, also known as the piotrowski test, is a biochemical assay that allows one to accurately quantify protein concentration within the range of 5150 mgml.
The bradford protein assay can measure protein quantities as little as 1 to 20. 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 bca protein assay 19 micro bca protein assay 20. Pierce protein biology thermo fisher scientific us. Instructions pierce 660nm protein assay 22660 22662 2070. Piercea protein assay kit bc thermo fisher scientific pdf. Pdf a comparison of protein quantitation assays for. Thermo scientific pierce protein molecular weight marker thermo scientific pierce prestained protein molecular weight markers are designed to monitor protein migration during sdspolyacrylamide gel electrophoreses and monitor protein transfer onto membranes. Pdf protein content quantification by bradford method. The amino acid backbone forms a colorchelate complex with the copper molecules, allowing for a highly sensitive and selective detection of the. A comparison of protein quantitation assays for biopharmaceutical applications article pdf available in molecular biotechnology 372. When completed according to the pierce 660nm protein assay protocol, protein detection is linear on the glomax discover. If precise quantitation of an unknown protein is required, it is advisable to select a protein. The dye reagent concentrate can be purchased in a kit with one of two standards.
The assay is more linear than coomassiebased bradford assays and compatible with higher concentrations of most detergents, reducing agents, and other commonly. Thermo scientific pierce quantitative colorimetric peptide. The reagent to sample ratio used for the pierce 660 nm protein assay was 15. The pierce 660 nm protein assay has a greater linear range of 252,000. A read is counted each time someone views a publication summary such as the title, abstract, and list of authors, clicks on a figure, or views or downloads the fulltext. 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. 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. Thermo scientific pierce bca protein assay kit remains one of the most popular protein quantitation methods worldwide. The pierce bca assay can be used to assess yields in whole cell. Bradford dye is easy to use, as well as fast and sensitive, but several compounds can interfere with the assay see the pdf file for the biorad protein assay. Qualitative test for protein indicates the type of protein and is done by chemical methods. Pierce bca protein assay kit university of arizona. Then it is also done in quantitative terms whereby the amount of protein in a given biological sample is known. Bovine gamma globulin kit i, catalog number 5000001 or bovine serum albumin kit ii, catalog number 5000002.
All colorimetric protein assays require protein standard to estimate the concentration of a sample. Thermo scientific pierce 660 nm protein assay reagent. Pierce bca protein assay kit thermo fisher scientific. The total time it takes to set up and complete the assay is under 30 minutes. Protein assay reagents involve either protein dye binding chemistry coomassiebradford or protein copper chelation chemistry. Pierce bca protein assay kit, sufficient reagents for 500 testtube or 5000 microplate.
In this reaction, the copper is first reduced by the amide backbone of peptides under alkaline conditions biuret reaction. The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Researchers look to us for a range of innovative, highquality thermo scientific products, including our pierce antibody collection, supersignal chemiluminescent substrates, bca and other protein assay and analysis reagents, slidealyzer dialysis cassettes, cell lysis reagents, mass spectrometry sample prep and quantitation reagents, western blotting reagents and instruments, crosslinking and. When protein binds, the pka of the dye shifts causing the dye to become blue. An adaptation of pierces bca protein assay reagent kit product no. Pierce bca protein assay kit is a twocomponent, highprecision, detergent compatible. Third, the bradford assay is not affected by many of the compounds that limit the application of the lowry assay. Studies of the bca assay for determination of total protein. Biochemical analysis of proteins relies on accurate quantitation of protein concentration. However, because this method is laborious and typically yields.
Bca protein assay introduction the glomaxmulti jr and the pierce bca protein assay provide a simple method for quantifying protein over a large range. Pierce prediluted bsa standards were used to generate concentration response curves for both the pierce 660 nm assay and bradford assay on the nanodrop 8000. Measuring protein concentration using the 660nm protein assay. Pierce offers seven colorimetric assays for detection and quantitation of total protein. 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 is more linear than coomassiebased bradford assays and compatible with higher concentrations of most det. Firefighting measures promptly isolate the scene by removing all persons from the vicinity of the incident if there is a fire. The glomax discover system and pierce bca protein assay provide a simple method for quantifying protein over a large range.
Without protein, the solution is redbrown in its acidic solution. Jul 16, 2015 bradford assay principle explanation this lecture explains about the bradford assay to determine the total protein concentration in a cell. The primary focus of the unit is assay selection, emphasizing sample and buffer compatibility. Hazardous thermal decomposition products specific hazards arising from the chemical. The thermo scientific pierce 660nm protein assay is a quick, readytouse colorimetric method for total protein quantitation. This was done by analyzing the precision, accuracy, linearity and range of the method. Assays were performed according to the standard testtube procedure using 100. Pierce protein assays provide a wide range of options for accurate protein concentration determination based on assay time, sensitivity, compatibility, standard curve linearity, and protein to. The bradford protein assay is based on the observation that the absorbance maximum for an acidic solution of coomassie brilliant blue g250 shifts from 465 to 595 nm when binding to protein occurs. It provides quick estimation of the protein amount in a given sample. The bca assay is a twostep colorimetric assay used to quantitate the total protein in a sample. Thermo scientific pierce protein assay technical handbook.
Pierce bca reagents provide accurate determination of protein concentration with most sample. Piercea protein assay kit bc thermo fisher scientific. Mar 27, 2020 prepared from the protein and assayed alongside the unknowns before the concentration of each unknown is determined based on the standard curve. Pierce bca protein assay kit is a twocomponent, highprecision, detergentcompatible protein assay for determination of protein concentration. Using the glomax discover system with the pierce 660nm protein assay provides a convenient procedure for quantifying protein, and the assay protocol comes preloaded on the instrument listed as bradford assay. Thermo scientific pierce bca protein assay kit fisher. Streamline bcabased protein quantitation molecular devices. Proteintoprotein variation 6 references 7 introduction the pierce micro bca protein assay reagent kit is a detergentcompatible bicinchoninic acid formulation for the colorimetric detection and quantitation of total protein. Bradford protein assay an overview sciencedirect topics. Protein assays based on these methods are divided into two categories. Thermo scientific pierce protein molecular weight marker.
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