Thermo scientific pierce protein a, g, ag and l coated plates provide alternatives to direct, passive adsorption methods for immobilizing antibodies for elisa and other platebased assay techniques. Thermo scientific pierce 660 nm protein assay reagent. Bca is a highly sensitive and selective detection reagent for the. Protein interaction technical handbook 1601666 areas of interest. Table of contents introduction 17 enzymelinked immunosorbent assays elisas 1. 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. The actual concentrations for sample a and b are calculated by the linear equation obtain in the figure 1. Gel electrophoresis is a technique in which charged molecules, such as protein or dna, are separated according to physical properties as they are forced through a gel by an electrical current. Quick technical summaries pierce protein and peptide assays. Thermo scientific pierce protein assay technical handbook, 2015 your bibliography. Pierce bca protein assay kit pdf book manual free download. Thermo scientific pierce assay development technical handbook version 2. The reagent to sample ratio used for the pierce 660 nm protein assay was 15.
Selection of the protein assay when it is necessary to determine the total protein concentration in a sample, one of the first factors to consider is the selection of a protein assay method. It is important to evaluate the compatibility of each assay with sample types, assay range, sample volume, and the availability of a suitable spectrophotometer, as well as the time and cost. Protein yield comparison of two bacterial cell lysis reagents. On the basis of these findings, we developed a very.
The bicinchoninic acid assay bca, also referred to as the smiths assay, is a biochemical assay designed by paul smith in 1985. The pierce bca assay can be used to assess yields in whole cell. Read online pierce bca protein assay kit book pdf free download link book now. 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. Pierce bca protein assay kit university of arizona. Ripa lysis and extraction buffer thermo scientific ripa lysis and ripa lysis and extraction buffer protein extraction.
The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Protein assay compatibility notes pierce bca assay and coomassie plus assay protease inhibitors3 may be added to prevent protein degradation. Protein assay reagents involve either proteindye binding chemistry coomassiebradford or proteincopper chelation chemistry. The assay is very simple and quick to perform and is compatible with reducing agents and some detergents. Calculation of results 12 thermo scientific pierce 660nm protein assay overview highlights typical response curves 14 bcabased protein assays 15. Pierce offers seven colorimetric assays for detection and quantitation of total protein. Pierce chemical technical library bicinchoninic acid bca method one of the most recent advances in protein assay technology is the use of bicinchoninic acid bca in a unique reagent system. Table 1 summarizes the common total protein quantitation assays.
Protein yields micrograms for membrane, cytosolic, and total fractions were determined with the thermo scient ific pierce bca protein assay kit cat. The bicinchoninic acid assay, also known as the smith assay, after its inventor, paul k. If precise quantitation of an unknown protein is required, it is advisable to select a protein. Table of contents total protein assays quick technical summaries 1 introduction 4 selection of the protein assay 4. The choice among available protein assays is usually based. Protein transfer technical handbook protein ladder considerations kda 200 typically, protein ladders or standards are included during gel electrophoresis for protein sizing after separation. Protein assay technical handbook 16 september 2009. Thermo scientific pierce protein assay technical handbook version 2. Conclusion 6 the bradford protein assay is used to. Assays were performed according to the standard testtube procedure using 100. For more information or to view additional products.
In this reaction, the copper is first reduced by the amide backbone of peptides under alkaline conditions biuret reaction. At boster, we are working on our detailed sample preparation protocols that cover more than 20 sample types and expecting to update this handbook in the near future. Pierce bca protein assay kit is a twocomponent, highprecision, detergentcompatible protein assay for determination of protein concentration. 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 assay development technical. Thermo scientific pierce protein a, g, and l coated plates. 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. Mechanism of coomassie brilliant blue g250 binding to. You must order using the pr suffix to get the discount pricing. Piercea protein assay kit bc thermo fisher scientific.
Purify gstfusion proteins with thermo scientific pierce. Thermo scientific pierce electrophoresis technical handbook featuring thermo scientific gelcode staining kits. Red 660 protein assay offers greater linearity, greater color stability, and greater compat ibility with detergents, reducing agents and other interfering agents. In the part 1 of the experiment, the standard curve of absorbance versus concentration of bsa is constructed. Table of contents introduction 17 enzymelinked immunosorbent assays elisas 1 typical elisa protocol summary 4. Pierce bca reagents provide accurate determination of protein concentration with most sample types encountered in protein research. Download the protein assay technical handbook to help select the appropriate assay method based on the assay time, sensitivity, compatibility, standard curve linearity, and proteinto protein variation. Red 660 protein assay is a single reagent colorimetric assay that outperforms commercial colorimetric assays, including bradford and improved coomassie bradford assays. Calculation of results 12 thermo scientific pierce 660 nm protein assay overview highlights typical response curves 14 bcabased protein assays 15 chemistry of the bca protein assay 15. Bca protein assay kit from thermo scientific pierce. Thermo scientific pierce protein assay technical handbook. As shown in the figure 1, the absorbance reading increases with the concentration of bsa increases. Pierce protein biology thermo fisher scientific us. The bradford protein assay is used to estimate the concentration of protein.
Pierce assay has a significantly greater linear dynamic range than the bradford assay. Outside the united states, contact your local branch office or distributor. Thermo scientific pierce electrophoresis technical handbook. They are all wellcharacterized, robust assays that provide consistent, reliable results. All books are in clear copy here, and all files are secure so dont worry about it. Summary of contents for thermo scientific nanodrop one. This site is like a library, you could find million book here by using search box in the. We investigated the mechanism of coomassie brilliant blue g250 cbb binding to proteins in order to develop a protein assay with the maximum possible sensitivity. Thermo scientific pierce bca protein assay kit bca. Bradford protein assay wikimili, the best wikipedia reader. Follow the assay kit manufacturers recommendations for all standards and samples unknowns. 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. Votre bibliographie chemistry bibliographies cite this. Protein a, g and ag coated plates 12 protein l coated plates.
Thermo scientific pierce protein assay technical handbook thermo fisher scientific firm. Stoscheck cm 1987 protein assay sensitive at nanogram levels. Protein assay reagents involve either proteindye binding chemistry coomassie bradford or proteincopper chelation chemistry. Protein assay versus the thermo scientific pierce 660 nm protein assay. Page 94 protein assay kits and protocols please refer to the nanodrop website for uptodate kits and protocols for the nanodrop one instruments. Salts, chelating agents and reducing agents can be added for more efficient lysis. Thermo scientific pierce immobilized glutathione agarose recover gstfusion proteins for 30% less.
They are all wellcharacterized, robust assays that. Table of contents total protein assays quick technical summaries 1 introduction 4. We found that the neutral ionic species of cbb binds to proteins by a combination of hydrophobic interactions and heteropolar bonding with basic amino acids. Protein assay for technical service call your local biorad office or in the u. Compared to most dyebinding methods, the bca assay is affected much less by protein compositional differences, providing greater proteintoprotein uniformity. Protein yields micrograms for membrane, cytosolic, and total fractions were determined with the thermo sci entific pierce bca protein assay kit cat. The total protein concentration is exhibited by a color.
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