Signal detection may become saturated when measuring very high lightemitting samples in a luminometer or scintillation counter. The dual luciferase reporter dlr assay system provides an efficient means of performing dualreporter assays. For luciferase activity, pipet 20ul of each lysate into a 5ml polystyrene falcon tube. The celltiterglo luminescent cell viability assay is a homogeneous method to determine the number of viable cells in culture based on quantitation of the atp present, which signals the presence of metabolically active cells. The approximately 2hour signal halflife provides greater flexibility in assay design. The luciferase assay system is a homogeneous assay system. For an overview and information on the development of the dualglo luciferase assay system, see sections 6. Systems contain one vial each of brightglo luciferase assay substrate and buffer. The luciferase assay system also can be used for quantitation in plant and bacterial cells, but only cell culture lysis reagent is. Promega luciferase assay systems an extremely sensitive reagent for rapid quantitation of firefly luciferase. Guidelines for transfecting luciferase reporter vector with fugene 6. The luciferase assay system is an extremely sensitive and rapid reagent for quantitation of firefly luciferase. The webinar also discusses the concept of the coincidence reporter assay, an assay developed at the nih that is especially useful for monitoring reporter activity due to promoter activation in screening applications.
Read on luminometer in the gel room using promega luciferase assay substrate thawed and mixed at rt. Other than comparable commercial reporter assay systems and their noncommercial counterparts, the prluc assay system was formulated under the aspect of full compatibility with standard methods for protein. Multiplexing luciferase reporter and cell viability assays. The enliten atp assay system is used to measure atp levels for the indirect detection of. The thiol reagents other than coa are reagents which have a free sulfhydryl group that is capable of being effective as a reducing agent in an airsaturated aqueous solution under conditions, of temperature, ph, ionic strength, chemical composition, and. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 6082744330 fax 6082772516. Email promega technical services if you have questions on use of this system. Cypridina is a new secreted luciferase reporter from the ostracod cypridina noctiluca. When describing a procedure for publication using this product, we would appreciate it if you would refer to it as the luciferase reporter vector from panomics if a paper cites the luciferase reporter vector and is published in a research journal, the lead authors may receive a travel stipend for use at a. Although the assay can be performed effectively with solutions prepared in the laboratory, the commercially available luciferase reporter vectors and assay kits provide excellent linear response over. Among the assay systems, in which chemiluminescence has been employed to monitor or measure events of interest, are those in which the activity of a bioluminescent enzyme, a luciferase, is measured. The celltiterglo one solution assay is a homogeneous method of determining the number of. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.
Optimization of the luciferase assay kit may be necessary for use with your equipment or samples. It is widely used because it is convenient, relatively inexpensive, and gives quantitative measurements instantaneously. We established a quantitative reporter gene protocol, the prluc assay system, allowing the sequential measurement of photinus and renilla luciferase activities from the same extract. Total elapsed time for the dlrz assay 30 seconds promega corporation.
Now that you have a better understanding of how these two products differ, i want to mention one more option for consideration in the quest to find your best fit dualluciferase assay system. Enliten atp assay system bioluminescence detection kit for atp measurement technical bulletinpdf 427 kb english. Systems include stop and glo buffer and substrate, one vial of luciferase assay substrate, 10 ml luciferase assay buffer ii, and 30 ml passive lysis 5x buffer. Endpoint assays can provide sensitive, highthroughputamenable assay formats for determining cell health. Luciferase assay system, 10pack e1501 from promega. I am carrying out the dual luciferase assay promega on different cell lines. General considerations the steadyglo luciferase assay system is designed for use with the following culture media containing 010% serum. Cell viability and cytotoxicity assays are based on colorimetric, fluorometric and bioluminescent detection chemistries. This amount of cells should yield 5080% confluence on the day of transfection. The luciferase assay system is substantially improved over conventional assay methods in both sensitivity and simplicity. For kits e1500, e1501, e4030, e4530 and e4550, prepare luciferase assay reagent lar by adding luciferase assay buffer 10ml for e152a and 100ml for e152b to the vial of lyophilized luciferase assay substrate. The coincidence reporter system is designed to identify false hits more rapidly so time and money are not wasted.
Luciferase assay systems instructions for use of products e1483, e1500, e1501, e1531, e4030, e4530 and e4550. E3971 can be used with the luciferase assay system for reporter quantitation in mammalian cells. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. Feb 01, 1994 reporter molecules or labels in such assay systems have included radioactive isotopes, fluorescent agents, enzymes and chemiluminescent agents. The dualglo luciferase reagent can be added directly to cells in growth medium without washing or preconditioning. A search for dualluciferase on highwire press generates over 700 articles from 2009 alone. Usa toll free in usa 8003569526 6082744330 fax 6082772516.
Luciferase assay system 100 assays e1500 each system contains sufficient reagents for 100 standard assays. A search for dual luciferase on highwire press generates over 700 articles from 2009 alone. Ideal for high and ultrahighthroughput applications. Assay lysates for luciferase activity and protein concentration. Promega s multiplexed luciferase reporter and cell.
General considerations the nanoglo dualluciferase reporter assay system is designed to be used with many media types and has been validated for use with the following culture media containing 010% serum. Dec 21, 2010 more importantly, as we have shown here in fig. The dualglo luciferase assay system is designed to work in growth media commonly used for mammalian cells with or without added serum. Dualglo luciferase assay system quick protocol fbpdf 125 kb english. This luciferase does not require atp and catalyzes the oxidation of its luciferin substrate in a photochemical reaction. Transfection for 6well plate using genejuice transfection reagent requires 1. Celltiterglo one solution assay technical bulletinpdf. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526.
Can be used for assays in singletube luminometers or in multiwellplate luminometers with autoinjectors. Promega corp original assignee promega corp priority date the priority date is an assumption and is not a legal conclusion. We offer luciferase reporter reagents, multiple cloning site vectors, and flash, glow and dualspectral luciferase assay kits based on the intracellularly expressed firefly luciferase. Dual luciferase assay system for rapid assessment of gene. Promega corporation promega maxwell rsc viral total nucleic acid purification kit, 48 preps the maxwell rsc viral total nucleic acid purification kit is designed for automated extraction of viral total nucleic acid rna and dna from serum, plasma or other samples using the maxwell rsc instrument. Luciferase reporter assay kit university of michigan. Single luciferase reporter assay molecular cloning. All reagents were prepared as described by the manufacturer. The substrate for cypridina is different than coelenterazine, which.
Promega luciferase assay systems fisher scientific. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516. Luciferase based assays have become an invaluable tool for the analysis of cloned promoter dna fragments, both for verifying the ability of a potential promoter fragment to drive the expression of a luciferase reporter gene in various cellular contexts, and for dissecting binding elements in the promoter. To see articles that cite the use of the dualglo luciferase assay system, visit. Product components and storage conditions product size cat. Nih3t3 cells expressing the luciferase gene from rous sarcoma virus were lysed with 1x cell culture lysis reagent 48 hours after infection. Luciferase reporter assays allow you to investigate promoter activity by measuring light output from luciferase enzyme that is expressed under the control of your promoter of interest. The shaded area represents the light typically lost in measurements. Celltiterglo one solution assay promega corporation. To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. So why are dual luciferase reporter assays so widely.
Pdf dual luminescencebased reporter gene assay for. Download any file posted by another user of a community feature that you know, or reasonably should know, cannot be legally distributed in such manner or that you have a contractual obligation to keep confidential. After 24 hr rest, cells were stimulated with lps il10 for 2 hr. A turnon splitluciferase sensor for the direct detection of. So why are dualluciferase reporter assays so widely.
Divide the luciferase substrateassay buffer into aliquots 1ml per tube, 10 tubes to avoid multiple freezethaw cycles store at 80c, if using for longer than 1 month. If this occurs, dilute your sample with 1x lysis buffer and repeat the assay. Oneglo ex luciferase assay system provides both the high sensitivity and longlived luminescence required to batchprocess multiple 96, 384 or 1,536well plates. The dualluciferase reporter dlr assay system provides an efficient.
Instructions for use of products g7570, g7571, g7572, g7573. It has broad applications across various fields of cell and molecular biology wherever you want to measure or track expression of a cloned gene. Promega corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, highvalue products for the life sciences. Dualluciferase reporter assay system promega corporation. Pdf a turnon splitluciferase sensor for the direct. The luciferase reporter assay is commonly used as a tool to study gene expression at the transcriptional level. Linear results are seen over at least 8 orders of magnitude of enzyme concentration, and the patented technology incorporated into this assay system allows measurement of less than 1020 moles of luciferase under optimal conditions. Realtime, livecell assays repeatedly monitor over time and generate multiple data points from a single assay well. Firefly luciferase is a 60kda protein produced in nature by several species of the lampyridae family of beetles which includes the genera photinus and luciola. Light is produced by converting the chemical energy of luciferin oxidation through an electron transition, forming the product molecule oxyluciferin. The luciferase assay system yields linear results over at least eight orders of magnitude.
Apr 05, 2019 now that you have a better understanding of how these two products differ, i want to mention one more option for consideration in the quest to find your best fit dualluciferase assay system. Dualluciferase reporter assay system measures firefly and renilla luciferases from a single sample. Use biorad assay for protein concentrations in 20ul of. A firefly luciferase and b renilla luciferase signals were. As part of my job i occasionally search the literature for papers citing use of promega products in new or interesting ways. Promega luciferase assay system components fisher scientific. The dualluciferase reporter dlr assay system provides an efficient means of performing dualreporter assays. Comparison of the promega luciferase assay system to the conventional luciferase assay method. Linear results are seen over at least eight orders of magnitude of enzyme concentration, and less than 10 20 moles of luciferase can be measured under optimal conditions. Regain access you can regain access to a recent pay per article purchase if your access period has not yet expired. Dual luminescencebased reporter gene assay for luciferase and galactosidase. Cell culture lysis reagent and reporter lysis buffer may be used with the luciferase assay system for reporter quantitation in mammalian cells.
Performing the nanoglo dualluciferase reporter assay 3. Any search on dualluciferase reporters usually generates a lot of returns. Sep 01, 2006 such systems have found widespread application through the availability of commercial assay kits the dual luciferase reporter assay system, cat. In the dlr assay, the activities of firefly photinus pyralis and renilla renilla reniformis, also known as sea pansy luciferases are measured sequentially from a single sample. Luciferase assay to study the activity of a cloned promoter. The luciferase assay system incorporates coenzyme a coa for improved kinetics, allowing greater enzymatic turnover and resulting in increased light intensity that is nearly constant for at least 1 minute. The dualglo luciferase assay system is designed to allow highthroughput analysis of mammalian cells containing genes for firefly and renilla luciferases, grown in 96 or 384well plates. Cytotoxglo cytotoxicity assay technical bulletinpdf 1090 kb english. The behaviours of the transfected cells were evaluated by a fireflyluciferase reporter gene harbouring the promoter region of i. A protocol for combined photinus and renilla luciferase. The oneglo luciferase assay system provides a sensitive, robust, homogeneous assay for detection of firefly luciferase reporter gene expression in mammalian cells. Dualluciferase reporter assay system harvard university. Dualluciferase reporter assay system protocol promega. Bioluminescent atpbased assay for detecting live microbes in water.
Description the celltiterglo luminescent cell viability assayad is a homogeneous method to determine the number of viable. Waterglo system atp assay promega promega corporation. In the dlr assay, the activities of firefly and renilla luciferases are. Promegas luminescent celltiterglo assay 2 was multiplexed with either the luminescent enduren live cell substrate, or the luminescent viviren live cell substrate. For manual luminometers, the substrate solution is mixed by hand with the cell lysate, and the fluorescence is read at a defined time following mixing. Join researchgate to find the people and research you need to help your work. The celltiterglo one solution assayac provides a homogeneous method to. E1910, promega, madison, wi, usa and alternative protocols based on the same principles. Luciferase genes vary in stability, light output and spectral characteristics, and reporter kits are available with different luciferases for both single and dualspectral analyses. Generally, 100fold greater sensitivity can be achieved over the chloramphenicol acetyltransferase cat assay. Celltiterglo luminescent cell viability assay protocol.
Nanoglo dualluciferase reporter assay nanodlr system. The cytotoxglo assay is a luminescent cytotoxicity assay that measures the. The luciferase assay is extremely rapid, simple, relatively inexpensive, sensitive, and possesses a broad linear range. Addition of oneglo ex reagent lyses cells and provides the substrates to measure firefly luminescence. The biolux cypridina luciferase assay kit contains the reagents necessary for assaying cypridina luciferase activity. An extremely sensitive reagent for rapid quantitation of firefly luciferase. Firefly luciferase assays and vectors thermo fisher. Dual luciferase assays were performed with the dual luciferase reporter assay system promega corp. Any search on dual luciferase reporters usually generates a lot of returns. Our teams are in regular contact with suppliers and distributors worldwide and are taking all steps necessary to address both demands for diagnostic tools and reliable delivery of all products as quickly as possible.
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