In the dlr assay, the activities of firefly and renilla luciferases are measured sequentially from a single sample. E1910, promega, madison, wi, usa and alternative protocols based on the same principles. Dual luciferase reporter assay system promega e1910 optional. Dualluciferase reporter assay and dualluciferase reporter assay system quick protocol fb024 pdf 73. The dualluciferase reporter dlr assay system provides an efficient means of performing dualreporter assays. Reporter assay and dualluciferaser promega corporation. Promega dual luciferase reporter dlr assay systems the dualluciferase reporter dlr assay system provides an efficient means of performing two reporter assays. Protocol for dna transfection and luciferase assay using.
Using the synergyht multidetection microplate reader to. Here, we present a rapid and sensitive assay based on the classical dualluciferase reporter technique which can be used as a new tool to characterize the minimum promoter region of a gene as well as the in vivo response of inducible promoters to different stimuli. The convenient addmixmeasure protocol generates both firefly and renilla luciferase luminescence signals from cells that have not been preconditioned or. This firefly luciferase assay kit is designed for simple and efficient quantitation of firefly luciferase reporter enzyme activity from cultured cells with high sensitivity and linearity. The dual glo 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. Dualluciferase assay system pn e1910 and 5x luciferase passive lysis buffer pn e1941 were purchased from promega corporation madison, wi. The dualluciferase assay has been widely used in cell lines to determine rapidly but accurately the activity of a given promoter. A noncommercial dual luciferase enzyme assay system for.
Dual luciferase reporter assay and dual luciferase reporter assay system quick protocol fb024 pdf 73 kb english. The dual glo luciferase reagent can be added directly to cells in growth medium without washing or preconditioning. I performed a dual luciferase assay using pgl3 firefly luc as experimental reporter and prltk renilla luc as internal control promega assay kit. Dualluciferase reporter assay system protocol promega. This kit is a new and improved replacement for the firefly luciferase assay kit catalog no. The dualluciferase reporter dlr assay system provides an efficient means of performing two reporter assays. Assay cell lysates at desired time after transfection for the luciferase expression using dualluciferase assay system promega according to manufacturers instructions. The dualluciferase reporter dlr assay system provides an efficient means. Promega dualluciferase reporter dlr assay systems dual.
In the dlr assay, the activities of firefly and renilla luciferases are measured sequentially from a. The dual luciferase reporter dlr assay system provides an efficient means of performing two reporter assays. Supplier page from promega for dualluciferase reporter assay system 10pack. Technical manual dual luciferase reporter assay system instructions for use of products e1910 and e1960. Measuring the dualluciferase reporter assay on the glomax. Do not wash 293t cells since they detach easily place 200. We illustrate the usefulness of this system for studying both constitutive telomerase and inducible nf. Instructions for use of products e2920, e2940, e2980. For best results with the dualluciferase assay, promega recommends using a luminometer that has been validated for use with the assay.
Dualluciferase reporter assay system 10pack from promega. Dualluciferase reporter assay and dualluciferase reporter assay system quick protocol fb024 pdf 73 kb english. The nanoglo dual luciferase reporter nanodlr assay detects the activities of dual luciferases, firefly and nanoluc reporter enzymes, in a single sample. The dlr and dualglo assays provide rapid and convenient means for achieving greater control over the biological significance of.
Orders of magnitude greater sensitivity for nluc in nanodlr assay compared to renilla. After using the dual luciferase assay from promega, however, i am getting luminescence values from 300 to 1200 for firefly luciferase and 1530 for renilla luciferase background is 12. The dual luciferase reporter dlr assay systemac provides an efficient. These dual assay kits use pairs of luciferase enzymes that are spectrally resolved i. Improved reagent stability and signal stability to facilitate batch processing of plates 5.
The nanoglo dual luciferase reporter nanodlr assay system 4. The chromaglo luciferase assay system and the chromaluc vectors provide a method to generate red and green dualcolor luminescence from a single sample upon a singlereagent addition. Here, we present a rapid and sensitive assay based on the classical dualluciferase reporter technique which. Fast, sensitive detection of renilla luciferase activity. In the dlr assay, the activities of firefly photinus pyralis and renilla renilla reniformis or sea pansy luciferases are measured sequentially from a single sample. As part of my job i occasionally search the literature for papers citing use of promega products in new or interesting ways. Nanoglo dualluciferase reporter assay system promega. The dual luciferase reporter dlr assay system provides an efficient means of performing dual reporter assays. 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. Dualluciferase reporter assay system harvard university. Luciferase genes vary in stability, light output and spectral characteristics, and reporter kits are available with different luciferases for both single and dualspectral analyses. Any search on dualluciferase reporters usually generates a lot of returns. A search for dualluciferase on highwire press generates over 700 articles from 2009 alone. However, the assay is designed to allow for higher throughput processes given a considerably more stable luminescent signal for both reporters and homogenous assay format with no washing or preconditioning required.
Application of the dualluciferase reporter assay to the. Consecutive detection of firefly and renilla luciferase activities in a single sample. Cells, transfected with the plasmid of interest and a luciferase reporter. Dualluciferase reporter assay system e1910 from promega. The luciferase assay system was developed for reporter quantitation in mammalian cells. The components of the dlr assay system are identical in formulation to those provided with the dlr assay system cat. The thermo scientific pierce luciferase dual assays deliver high sensitivity in a simple onestep detection protocol.
A protocol for combined photinus and renilla luciferase. Dual luciferase reporter assay chemistry continued in the dlr assay chemistry, the kinetics of the renilla luciferase reaction provide a stabilized luminescent signal that. This system assays cells directly in growth medium. Dualluciferase reporter assay system technical manualpdf.
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. Although this strategy has proved very useful, it does not allow the promoter and gene function to be analyzed in the context of the whole organism. The firefly luciferase reporter is measured first by adding luciferase assay reagent ii lar ii to generate. Weve made big improvements to our luciferase assay kits, with simplified assay setup, improved working solution stability, and high sensitivity and linearity. So why are dualluciferase reporter assays so widely. Dualluciferase reporter assay system promega corporation. Luciferase assay system protocol promega corporation. Such systems have found widespread application through the availability of commercial assay kits the dualluciferase reporter assay system, cat.
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