Thermo scientific pierce western blotting handbook and troubleshooting guide in the united states. Thermo scientific pierce bsa protein digest standard, lcms grade rt. Thermo scientific pierce mammalian galactosidase assay. The micro bca protein assay kit is a specialized version of the popular pierce bca protein assay for determining the protein concentration of dilute samples. Search for library items search for lists search for contacts search for a library. Protein concentration was determined using a pierce bca protein assay kit thermo fisher scientific, 23227. Thermo scientific pierce electrophoresis technical handbook. Thermo scientific evolution 260 bio spectrophotometer. Quick technical summaries pierce protein and peptide assays. Thermo scientific pierce protein molecular weight marker. This method has a fast detection speed and low proteintoprotein variability in comparison to the bca or coomassie bradford. Pierce protein ladders many available, see pages 712. It is a popular protein assay, due to its compatibility with detergents.
Mercury lamp calibration accessory calibration validation carousels general, ep, usp. Figures 35, shows the low cvs of the standards, indicating consistent sample preparation across. Protein yields micrograms for membrane, cytosolic, and total fractions were determined with the thermo sci entific pierce bca protein assay kit cat. Protein identification was performed with thermo scientific proteome discoverer software and the ipi protein database, and relative peptide quantification was performed simultaneously by comparing the abundance ratios of the reporter ions from the tmt tags. Intelligent microvolume analysis thermo fisher scientific. Pierce fenta phosphopeptide enrichment kit 5354 cell surface protein isolation kit 55 glycoprotein isolation kits 56 ubiquitin enrichment kit 57 antibody purification 5867 overview 58 immobilized protein l, protein a, protein g 5961 and protein ag igg binding and elution buffers for 62 protein a, g, ag and l. Piercea protein assay kit bc thermo fisher scientific. Nucleic acid interactions 32 crosslinking agents 33 related crosslinking products 35 nucleic acidbiotin conjugates 36. Thermo scientific pageruler unstained protein ladders thermo scientific pageruler unstained protein ladders are mixtures of recombinant, highly purified proteins designed for accurate sizing of proteins by sdspage.
Protein assay the wolfson centre for applied structural biology wolfson. Thermo scientific pierce 660 nm protein assay reagent. For more information or to view additional products. We purchased the pierce bca protein assay kit thermo scientific with bsa standards. Thermo scientific evolution 260 bio spectrophotometer brochure. Shop online for a wide selection of thermo scientific pierce mammalian galactosidase assay kit lyse cultured mammalian. Thermo scientific pierce electrophoresis technical handbook featuring thermo scientific gelcode staining kits version 2 table of contents thermo scientific pierce products for gel electrophoresis of proteins gel electrophoresis is a technique in which charged molecules, such as protein or dna, are separated according to physical properties as. Electrophoresis technical handbook thermo scientific. The assay is very simple and quick to perform and is compatible with reducing agents and some detergents. Protein assay versus the thermo scientific pierce 660 nm protein assay. Protein assay technical handbook thermo fisher scientific.
Bovine gamma globulin kit i, catalog number 5000001 or bovine serum albumin kit ii, catalog number 5000002. Dilute the contents of one albumin standard bsa ampule. Jun 15, 2011 thermo scientific pierce protein biology products are the cornerstone of the proteomics portfolio for thermo fisher scientific. This type of assay is called a sandwich assay because the analyte to be measured is bound between two antibodies the capture antibody and the detection antibody. Protein assay reagents involve either protein dye binding chemistry coomassiebradford or protein copper chelation chemistry. This method may be preferred for samples containing detergents or other reducing agents. Assays were performed according to the standard testtube procedure using 100. 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.
Thermo scientific pierce 660nm 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 protein assay reducing agent compatible 18 bca protein assay 19 micro bca protein assay 20. Thermo scientific pierce protein biology products youtube. Smad3 regulates neuropilin 2 transcription by binding to. Thermo scientific pierce microplate bca protein assay kit. Thermo scientific pierce protein purification technical handbook. Thermo scientific pierce bca protein assay kit protein. Pierce protein biology thermo fisher scientific us. This reducing agentcompatible rac version of the thermo scientific pierce bca protein assay provides accurate determination of protein concentration in samples that contain dithiothreitol dtt, 2mercaptoethanol bme, tcep and other disulfide reducing agents. Prepare samples and blanks basic instrument operations protein a280 reported results protein a280 measurement screen for each measured sample, this application shows the absorbance spectrum and a summary of the results.
Tools and reagents for improved quantitation of total or. Page 1 nanodrop microuvvis spectrophotometers nanodrop one user guide 269309101 revision b july 2016 page 2 telephone. The thermo scientific pierce bca protein assay kit. Protein assay compatibility table thermo fisher scientific. Thermo scientific halt and thermo scientific pierce protease and phosphatase inhibitor cocktails and tablets are broadspectrum blends in both liquid 100x and tablet formats for complete protein protection during extraction.
Mixing together the three micro bca reagents results in a working solution that is sufficiently concentrated to measure protein when. Thermo scientific pierce rapid gold bca protein assay kit is twocomponent, highprecision, detergentcompatible assay optimized to measure at 480nm total protein concentration compared to protein standard curve of known concentrations. Contents protein cleanup technical handbook introduction 2 protein dialysis using regenerated cellulose membranes overview 4 membrane performance data 6 selecting the right format for sample processing 8 pierce 96well microdialysis plate 9 slidealyzer mini dialysis devices 10 original thermo scientific slidealyzer dialysis cassette 11. Detergents are frequently used in cell lysis reagent formulation and other protein. Pierce protein research products po box 117 stain without pain. Thermo scientific pierce antibody production and purification technical. Simply dilute the 10x solution with water and use directly in tank or conventional semidry transfer. Votre bibliographie chemistry bibliographies cite this. This method combines the well known reduction of cu. To alleviate the frustration of discovering these incompatibilities by trial and error, our technical support scientists have characterized the effects of commonly used buffers and sample components on the behavior of seven different thermo scientific pierce protein assays. Detect and measure hydrogen peroxide levels in biological samples using an iron and xylenol orange reagent with thermo scientific pierce quantitative peroxide assay kits. Protein cleanup technical handbook the macromolecules e. Conclusion 6 the bradford protein assay is used to estimate. The sandwich format is used because it is sensitive and robust.
Protocol for bca protein assay using a nanodrop 20002000c spectrophotometer subject. Thermo scientific pierce bca protein assay kit from thermo. Pierce bca protein assay kit brigham young university. The recently launched thermo scientific pierce 660 nm protein assay eliminates many of the bradford assay issues that can occur, including problems associated with microvolume measurements. Votre bibliographie chemistry bibliographies cite this for me. If the total protein concentration in the samples is high 2,000. We offer a complete array of products to support rapid and ef. Thermo scientific pierce bca protein assay kit remains one of the most popular protein quantitation methods worldwide. Thermo scientific pierce protein assay technical handbook version 2.
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. The ladders resolve into clearly identifiable sharp bands when analyzed by sdspage and stained with coomassie blue or silver stain. Used in more labs than any other detergentcompatible protein assay, pierce bca reagents provide accurate determination of protein concentration with most sample types. Download the protein assay technical handbook pierce protein. For those samples that contain detergents, copperbased protein assays such as the thermo scientific pierce rapid gold. P rotein preparation handbook thermo fisher scientific. Thermo scientific pierce pierce rapid gold bca protein. Design, visualize and detect immunostaining fisher scientific. Technical and operational assistance training support services. We offer kits, reagents and services that enable researchers to. Purify gstfusion proteins with thermo scientific pierce. Provides high accuracy results in fraction of time. Protein purifi ation various methods are used to enrich or purify a protein of interest from other proteins and components in a crude cell lysate or other sample. Protein yield comparison of two bacterial cell lysis reagents.
As shown in the figure 1, the absorbance reading increases with the concentration of bsa increases. View 1601975antibodyproductionpurificationguide from biotechnol 001 at international islamic university malaysia. Download the protein assay technical handbook to help select the appropriate assay method based on the assay time, sensitivity, compatibility, standard curve linearity, and protein to protein variation. Protein transfer technical handbook thermo fisher scientific. The pierce 660 nm protein assay has a greater linear range of 252,000. The pierce 660 nm protein assay has several advantages, including a simple format, a stable endpoint, a.
These aqueous and lipidcompatible peroxide assay kits detect peroxides based on oxidation of ferrous to ferric ion in the presence of xylenol orange. 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. Thermo scientific pierce western blotting handbook and. Protein transfer technical handbook pierce methanolfree transfer buffer, 10x thermo scientific pierce methanolfree transfer buffer does not require cooling or the addition of methanol. Thermo scientific pierce protein assay technical handbook, 2015 your bibliography. The dye reagent concentrate can be purchased in a kit with one of two standards. Quick technical summaries thermo scientific protein assays. While bradford assays can be performed on nanodrop. Protein transfer is a vital step in western blotting involving the transfer of proteins separated in a gel by electrophoresis to a solid support matrix so that speci. The easytouse thermo scientific pierce protein concentrators contain. Ripa lysis and extraction buffer thermo scientific ripa lysis and ripa lysis and extraction buffer protein extraction. In the part 1 of the experiment, the standard curve of absorbance versus concentration of bsa is constructed. The pierce lowry assay catalogue number 23240 was used in and biorad laboratories lowry assay was used in and for protein quantitation.
Biorad dc protein assay is a modified version of the lowry assay. Thermo scientific nanodrop one user manual pdf download. Thermo scientific pierce antibodies are developed for a wide. Thermo scientific pierce bca protein assay kit fisher. Thermo scientific pierce quantitative peroxide assay kits. How to create your own affinity titer column kelly j. 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 products for gel electrophoresis of proteins 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. The thermo scientific pierce bca protein assay kit is a twocomponent, highprecision, detergentcompatible assay reagent set to measure a562nm total protein concentration compared to a protein standard.
The assay is much more convenient than having to make up your own solutions for a biuret or lowry assay. 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. Outside the united states, contact your local branch office or distributor. 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.
Mixing together the three micro bca reagents results in a working solution that is sufficiently concentrated to measure protein when mixed with an equal volume of sample. Cells were lysed in radioimmunoprecipitation assay buffer 50 mmoll tris, 150 mmoll nacl, 1% nonidet p. Pierce chip reagents and kits 25 transcription factor assays 2831 chemiluminescent transcription factor assay kits 29 thermo scientific protein. Thermo scientific pierce assay development technical. Thermo scientific pierce protein assay technical handbook thermo fisher scientific firm home. For buffer exchange diafiltration, the concentrated solution is diluted and concentrated multiple times until the desired state is achieved. The protein and cell analysis education site is a freeaccess destination where you can learn more about applications and techniques related to protein gel chemistries, western detection, mass spectrometry, flow cytometry, fluorescence imaging, high content imaging, antibodies and antibody labeling, multiplex immunoassays, and more. They are all wellcharacterized, robust assays that provide consistent, reliable results. Thermo scientific pierce protein assay technical handbook. Measuring protein concentration using the bca protein assay with. Compared to most dyebinding methods, the bca assay is affected much less by protein compositional differences, providing greater protein to protein uniformity. Procedia chemistry 6 2012 184 a 193 18766196 2012 elsevier b. For more information, or to download product instructions, visit.
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