Cypridina is a new secreted luciferase reporter from the ostracod cypridina noctiluca. Promega luciferase assay systems fisher scientific. Pdf dual luminescencebased reporter gene assay for. It has broad applications across various fields of cell and molecular biology wherever you want to measure or track expression of a cloned gene. Celltiterglo luminescent cell viability assay protocol. Firefly luciferase assays and vectors thermo fisher. Systems contain one vial each of brightglo luciferase assay substrate and buffer. A firefly luciferase and b renilla luciferase signals were. Product components and storage conditions product size cat.
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. Sep 01, 2006 such systems have found widespread application through the availability of commercial assay kits the dual luciferase reporter assay system, cat. 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. Transfection for 6well plate using genejuice transfection reagent requires 1. The experimental setup was similar for each assay combination. An extremely sensitive reagent for rapid quantitation of firefly luciferase.
The dualglo luciferase reagent can be added directly to cells in growth medium without washing or preconditioning. Can be used for assays in singletube luminometers or in multiwellplate luminometers with autoinjectors. In the dlr assay, the activities of firefly and renilla luciferases are. Promega corp original assignee promega corp priority date the priority date is an assumption and is not a legal conclusion. Promega s multiplexed luciferase reporter and cell viability. The cytotoxglo assay is a luminescent cytotoxicity assay that measures the. The approximately 2hour signal halflife provides greater flexibility in assay design. The behaviours of the transfected cells were evaluated by a fireflyluciferase reporter gene harbouring the promoter region of i. The shaded area represents the light typically lost in measurements. 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. The luciferase assay is extremely rapid, simple, relatively inexpensive, sensitive, and possesses a broad linear range. Luciferase assay system, 10pack e1501 from promega. The biolux cypridina luciferase assay kit contains the reagents necessary for assaying cypridina luciferase activity.
So why are dualluciferase reporter assays so widely. Promega corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, highvalue products for the life sciences. The luciferase assay system is an extremely sensitive and rapid reagent for quantitation of firefly luciferase. 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. A protocol for combined photinus and renilla luciferase. Generally, 100fold greater sensitivity can be achieved over the chloramphenicol acetyltransferase cat assay. 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. 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. All reagents were prepared as described by the manufacturer. Promega luciferase assay systems an extremely sensitive reagent for rapid quantitation of firefly luciferase.
General considerations the steadyglo luciferase assay system is designed for use with the following culture media containing 010% serum. Luciferase assay systems instructions for use of products e1483, e1500, e1501, e1531, e4030, e4530 and e4550. The celltiterglo one solution assay is a homogeneous method of determining the number of. 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. Regain access you can regain access to a recent pay per article purchase if your access period has not yet expired. The oneglo luciferase assay system provides a sensitive, robust, homogeneous assay for detection of firefly luciferase reporter gene expression in mammalian cells. Enliten atp assay system bioluminescence detection kit for atp measurement technical bulletinpdf 427 kb english. To see articles that cite the use of the dualglo luciferase assay system, visit. As part of my job i occasionally search the literature for papers citing use of promega products in new or interesting ways.
For an overview and information on the development of the dualglo luciferase assay system, see sections 6. Oneglo ex luciferase assay system provides both the high sensitivity and longlived luminescence required to batchprocess multiple 96, 384 or 1,536well plates. 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. 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.
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. 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. The coincidence reporter system is designed to identify false hits more rapidly so time and money are not wasted. Luciferase reporter assay kit university of michigan.
The dual luciferase reporter dlr assay system provides an efficient means of performing dualreporter assays. Optimization of the luciferase assay kit may be necessary for use with your equipment or samples. Instructions for use of products g7570, g7571, g7572, g7573. 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. 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. Light is produced by converting the chemical energy of luciferin oxidation through an electron transition, forming the product molecule oxyluciferin. Email promega technical services if you have questions on use of this system. 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. Pdf a turnon splitluciferase sensor for the direct. The signal halflife under these conditions is greater than 5. Endpoint assays can provide sensitive, highthroughputamenable assay formats for determining cell health. Cell viability and cytotoxicity assays are based on colorimetric, fluorometric and bioluminescent detection chemistries. Dual luciferase assays were performed with the dual luciferase reporter assay system promega corp. A search for dual luciferase on highwire press generates over 700 articles from 2009 alone.
It is widely used because it is convenient, relatively inexpensive, and gives quantitative measurements instantaneously. Read on luminometer in the gel room using promega luciferase assay substrate thawed and mixed at rt. Luciferase assay system 100 assays e1500 each system contains sufficient reagents for 100 standard assays. 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. Addition of oneglo ex reagent lyses cells and provides the substrates to measure firefly luminescence. The luciferase assay system is substantially improved over conventional assay methods in both sensitivity and simplicity. Usa toll free in usa 8003569526 6082744330 fax 6082772516.
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. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed. The luciferase assay system also can be used for quantitation in plant and bacterial cells, but only cell culture lysis reagent is. Dualluciferase reporter assay system measures firefly and renilla luciferases from a single sample. Promegas luminescent celltiterglo assay 2 was multiplexed with either the luminescent enduren live cell substrate, or the luminescent viviren live cell substrate. To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. E3971 can be used with the luciferase assay system for reporter quantitation in mammalian cells.
Promega s multiplexed luciferase reporter and cell. Signal detection may become saturated when measuring very high lightemitting samples in a luminometer or scintillation counter. After 24 hr rest, cells were stimulated with lps il10 for 2 hr. The dualluciferase reporter dlr assay system provides an efficient means of performing dualreporter assays. Any search on dual luciferase reporters usually generates a lot of returns. Luciferase genes vary in stability, light output and spectral characteristics, and reporter kits are available with different luciferases for both single and dualspectral analyses. The luciferase assay system is a homogeneous assay system. Feb 01, 1994 reporter molecules or labels in such assay systems have included radioactive isotopes, fluorescent agents, enzymes and chemiluminescent agents. Realtime, livecell assays repeatedly monitor over time and generate multiple data points from a single assay well. Dualglo luciferase assay system quick protocol fbpdf 125 kb english. The enliten atp assay system is used to measure atp levels for the indirect detection of. Dec 21, 2010 more importantly, as we have shown here in fig.
Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 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. 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. The luciferase reporter assay is commonly used as a tool to study gene expression at the transcriptional level. 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. Join researchgate to find the people and research you need to help your work.
This luciferase does not require atp and catalyzes the oxidation of its luciferin substrate in a photochemical reaction. We offer luciferase reporter reagents, multiple cloning site vectors, and flash, glow and dualspectral luciferase assay kits based on the intracellularly expressed firefly luciferase. 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. Nih3t3 cells expressing the luciferase gene from rous sarcoma virus were lysed with 1x cell culture lysis reagent 48 hours after infection. Dualluciferase reporter assay system promega corporation.
Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526. Ideal for high and ultrahighthroughput applications. Performing the nanoglo dualluciferase reporter assay 3. Any search on dualluciferase reporters usually generates a lot of returns. The celltiterglo one solution assayac provides a homogeneous method to.
The dualglo luciferase assay system is designed to work in growth media commonly used for mammalian cells with or without added serum. Guidelines for transfecting luciferase reporter vector with fugene 6. So why are dual luciferase reporter assays so widely. A turnon splitluciferase sensor for the direct detection of. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516. Total elapsed time for the dlrz assay 30 seconds promega corporation. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. Comparison of the promega luciferase assay system to the conventional luciferase assay method.
For luciferase activity, pipet 20ul of each lysate into a 5ml polystyrene falcon tube. A search for dualluciferase on highwire press generates over 700 articles from 2009 alone. 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. Dual luciferase assay system for rapid assessment of gene. If this occurs, dilute your sample with 1x lysis buffer and repeat the assay.
Multiplexing luciferase reporter and cell viability assays. Single luciferase reporter assay molecular cloning. The luciferase assay system yields linear results over at least eight orders of magnitude. Dualluciferase reporter assay system harvard university.
Use biorad assay for protein concentrations in 20ul of. I am carrying out the dual luciferase assay promega on different cell lines. 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. Celltiterglo one solution assay promega corporation. Celltiterglo one solution assay technical bulletinpdf. Nanoglo dualluciferase reporter assay nanodlr system. 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. Cytotoxglo cytotoxicity assay technical bulletinpdf 1090 kb english. 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. E1910, promega, madison, wi, usa and alternative protocols based on the same principles.
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. Waterglo system atp assay promega promega corporation. Dual luminescencebased reporter gene assay for luciferase and galactosidase. The substrate for cypridina is different than coelenterazine, which. Promega luciferase assay system components fisher scientific.
Dualluciferase reporter assay system protocol promega. Luciferase assay to study the activity of a cloned promoter. Assay lysates for luciferase activity and protein concentration. Cell culture lysis reagent and reporter lysis buffer may be used with the luciferase assay system for reporter quantitation in mammalian cells. The dualluciferase reporter dlr assay system provides an efficient. This amount of cells should yield 5080% confluence on the day of transfection.
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