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. The modifications enable protein concentration determination for concentrated lysates protein 2 mgml. Microplate bca assay for wholecell lysate entered by kevin janes. Pierce bca protein assay kit pdf book manual free download. Pierce bca protein assay kit reducing agent compatible. The enhanced protocol requires a 60c incubation for 30 minutes. Quick technical summaries pierce protein and peptide assays. This system has been used to study immune cells in a few published reports, yet a comprehensive protocol optimized particularly for lymphocytes is lacking. Quick technical summariespierce protein and peptide assays. Pdf bicinchoninic acid bca assay in low volume researchgate. Mar 27, 2020 download a modulus method for bca protein assay book pdf free download link or read online here in pdf. Has anyone successfully used a 595 nm filter for this assay. L of bsa in transparent, normal volume 384well plates and incubating at 37 c produced results with good linearity r 2 0. Bca protein assay kit reducing agent compatible k818.
Smith at the pierce chemical company in 1985 smith et al. This protocol makes use of thermo scientifics bca protein assay in a 96well tissue culture plate format to measure the amount of. Patient care protocols pierce county, wa official website. Microplate bca assay for wholecell lysate entered by kevin janes janes lab protocols 31209 1 1. The manufacturer of the bca kit advises to use a 562 nm filter, or at most a 590 nm filter. For more information, or to download product instructions, visit. Read online a modulus method for bca protein assay book pdf free download link book now.
Pierce bca protein assay kit university of arizona. Highthroughput quantitation of firefly photinus pyralis luciferase expression in mammalian cells is commonly performed by batch processing of 96 and 384well plates. 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. 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. Evaluation of colorimetric bcabased quantification of hydrazide. Pierce bca reagents provide accurate determination of protein concentration with most sample types encountered in protein research. The protein concentration can be determined with red660 protein assay cat.
Read online pierce bca protein assay kit book pdf free download link book now. Perform the assay as described in the protocol and multiply the protein concentration of the sample by 1. The bicinchoninic acid bca 2 assay is a colorimetric method that is commonly employed to estimate the concentration of protein in a sample. Compare all available bca protein assays features of the bca protein assay kitreducing. However, decreasing the total volume to between 8 and 20. Compared to most dyebinding methods, the bca assay is affected much less by protein compositional differences, providing greater proteintoprotein uniformity. Consistent with the pierce bca kit protocol, mixing 60. Highthroughput quantitation of firefly photinus pyralis luciferase expression in mammalian cells is commonly performed by batch processing of 96 and 384well plates steadyglo luciferase assay system is designed for this purposeproviding longlived luminescence when added to cultured cel. The kit enables you to measure protein concentration in samples that contain the disulfide reducing agents. 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. Microplate bca assay for wholecell lysate entered by. 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. Lowry protein assay an overview sciencedirect topics. The extracellular flux analyzer is a commonly used device for evaluating the performance of glycolysis and mitochondrial respiration in many cell types.
I recently migrated from the bradford method to the microbca method from pierce for the quantification of total protein from cell culture. Measuring protein concentration using the bca protein assay. 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 bicinchoninic acid bca assay, first described by smith et al. This protocol describes how codemark australia certification bodies cbs must assess products for conformity with the performance requirements of the bca under the codemark australia scheme rules. I used bsa provided in the kit as a standard and the correlation coefficient was very good 0. Codemark protocol for the assessment of products against.
Download pierce bca protein assay kit book pdf free download link or read online here in pdf. The concentration cannot be determined either by uv absorbance at 280nm or bca protein assays due to the presence of the nhs leaving group. Do duplicates of all samples including the standards. Bca enhanced protocol assay was performed on the modulus using the. This site is like a library, you could find million book here by using search box in the. I want to do a bca protein microplate assay pierce but our plate reader only has a 595 nm filter. Thermo scientific pierce protein assay technical handbook. The protein carbonyl levels were determined as described in the assay protocol. Apr062016 i want to do a bca protein microplate assay pierce but our plate reader only has a 595 nm filter. This special adaptation of the popular pierce bca protein assay method enables accurate protein concentration measurement for samples containing dtt, 2me or tcep.
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. The two assays are of similar sensitivity, but since bca is stable under alkali conditions, this assay has the advantage that it. Pierce bca protein assay kit thermo fisher scientific. It uses the same copperchelating technology as the wellknown traditional pierce bca protein assay and provides comparable accuracy bu.
Bradford protein assay protocol university of san diego. Pierce micro bca protein assay free download as pdf file. The bicinchoninic acid bca assay for protein quantitation. Protein assay compatibility table from our website. Piercea protein assay kit bc thermo fisher scientific. The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total. Rna 1 1 1 eu pdf free download studies of electrode reactions and coordination geometries of cui. 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. A modulus method for bca protein assay pdf book manual.
Thermo scientific pierce microplate bca protein assay kit. The standard protocol permits either a 30minute incubation at 37c or a 2 hourincubation at room temperature. Thermo scientific pierce bca protein assay reagent a. Pierce micro bca protein assay proteins buffer solution. The bicinchoninic acid assay bca assay, also known as the smith assay, after its inventor, paul k. Dashedline entry indicates that the material is incompatible with the assay. The thermo scientific pierce bca protein assay kit. Its objective is to ensure that certification decisions by cbs are undertaken in an appropriate and consistent way. Bicinchoninic acid bca assay in low volume sciencedirect. 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. Protocol the following are instructions for the microplate assay format of the pierce bca protein assay kit thermo scientific cat. This bca protein assay kit is a reducing agentcompatible version of our popular pierce bca protein assay. The pierce rapid gold bca protein assay kit is a fast, twocomponent, highprecision, detergentcompatible assay optimized to determine total protein concentration.
Several methods are available, each having features that suit it to a particular use. Lecture notes titile unit titile get lecture notes. We would like to show you a description here but the site wont allow us. Thermo scientific pierce protein assay technical handbook nugi. The bca protein assay is used for quantitation of total protein in a sample. All books are in clear copy here, and all files are secure so dont worry about it. In test tubes or 96well plates, the relationship between protein concentration and absorbance is nearly linear over a. Module and the bca protein assay is 5 gml figure 1 and the maximum is 2 mgml figure 2.
Download a modulus method for bca protein assay book pdf free download link or read online here in pdf. Prepare pierce bovine serum albumin bsa standards 2,000. The thermo scientific pierce bca protein assay kit is a detergent compatible formulation based on bicinchoninic acid bca for the. Measuring the protein concentration of samples is essential for many protein assays, such as western blotting or zymography. Pierce bca protein assay kit is a twocomponent, highprecision, detergentcompatible protein assay for determination of protein concentration. Tools and reagents for improved quantitation of total or. Bsa standard curves were generated by softmax pro software, and a quadratic curve fit was applied, as recommended in the assay kit user guides. We did the pierce bca assay and ran the standard curve and were able to estimate protein concentrations in our samples.
Add 10l of bsa standards stored in 80c freezer to each of the combine 20ml bca protein assay reagent a pierce. Streamline bcabased protein quantitation molecular devices. Available in tube and microplate formats, these reducing agent compatible rac bca kits extend the already broad reagent compatibility of the bca protein assay protocol to include. The bicinchoninic acid bca assay is a protein quantification technique that was first demonstrated by paul k. Thermo scientific pierce microplate bca protein assay. Preparation of standards and working reagent required for both assay procedures. The following protocol is designed for use with the kingfisher flex instrument. Micro bca protein determination modified protocol developed by anand asthagiri 41697 modified by kevin janes 121702 use the pierce bsa standard 2000 gml. The principle of this technique is quite similar to the lowry protein assay, which is one of the alternative protein quantification methods. The determination of protein concentration is frequently required in biochemical work. Read online protocol for bca protein assay using a nanodrop 20002000c.
The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. 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. Mobile internet protocol and transport layer download. How to calculate concentration using the microbca kit. 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. We had excess standards so we decided to save them and are currently. Pierce bca protein assay kit, sufficient reagents for 500 testtube or 5000. Thermo scientific pierce bca protein assay kit protein. This site is like a library, you could find million book here.
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