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. Orders of magnitude greater sensitivity for nluc in nanodlr assay compared to renilla. The nanoglo dual luciferase reporter nanodlr assay system 4. The dlr and dualglo assays provide rapid and convenient means for achieving greater control over the biological significance of. Dualluciferase reporter assay system protocol promega. Luciferase genes vary in stability, light output and spectral characteristics, and reporter kits are available with different luciferases for both single and dualspectral analyses. 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 dual luciferase reporter dlr assay system provides an efficient means of performing dual reporter assays. Improved firefly chemistry and the bright, sensitive nanoluc luciferase make this a highly sensitive dual luciferase assay that is simple to use, provides superior data quality. Dual luciferase reporter assay chemistry continued in the dlr assay chemistry, the kinetics of the renilla luciferase reaction provide a stabilized luminescent signal that. The firefly luciferase reporter is measured first by adding luciferase assay reagent ii lar ii to generate. Do not wash 293t cells since they detach easily place 200. In the dlr assay, the activities of firefly and renilla luciferases are measured sequentially from a.
The dualluciferase reporter dlr assay system provides an efficient means. Technical manual dual luciferase reporter assay system instructions for use of products e1910 and e1960. 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. Dual luciferase reporter assay system promega e1910 optional. Promega dual luciferase reporter dlr assay systems the dualluciferase reporter dlr assay system provides an efficient means of performing two reporter assays.
Instructions for use of products e2920, e2940, e2980. These dual assay kits use pairs of luciferase enzymes that are spectrally resolved i. The dual luciferase reporter dlr assay systemac provides an efficient. Nanoglo dualluciferase reporter assay system promega. The dualluciferase assay has been widely used in cell lines to determine rapidly but accurately the activity of a given promoter. The convenient addmixmeasure protocol generates both firefly and renilla luciferase luminescence signals from cells that have not been preconditioned or. Luciferase assay system protocol promega corporation. 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. 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. Dualluciferase reporter assay system promega corporation.
Dualluciferase reporter assay system e1910 from promega. Protocol for dna transfection and luciferase assay using. The dual glo luciferase reagent can be added directly to cells in growth medium without washing or preconditioning. A protocol for combined photinus and renilla luciferase. Assay cell lysates at desired time after transfection for the luciferase expression using dualluciferase assay system promega according to manufacturers instructions. I performed a dual luciferase assay using pgl3 firefly luc as experimental reporter and prltk renilla luc as internal control promega assay kit. 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. Weve made big improvements to our luciferase assay kits, with simplified assay setup, improved working solution stability, and high sensitivity and linearity.
For best results with the dualluciferase assay, promega recommends using a luminometer that has been validated for use with the assay. 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 luciferase reporter dlr assay system provides an efficient means of performing two reporter assays. As part of my job i occasionally search the literature for papers citing use of promega products in new or interesting ways. We illustrate the usefulness of this system for studying both constitutive telomerase and inducible nf. Supplier page from promega for dualluciferase reporter assay system 10pack. Measuring the dualluciferase reporter assay on the glomax. Dualluciferase reporter assay system 10pack from promega.
Dualluciferase reporter assay system technical manualpdf. A noncommercial dual luciferase enzyme assay system for. Cells, transfected with the plasmid of interest and a luciferase reporter. The nanoglo dual luciferase reporter nanodlr assay detects the activities of dual luciferases, firefly and nanoluc reporter enzymes, in a single sample. Dualluciferase assay system pn e1910 and 5x luciferase passive lysis buffer pn e1941 were purchased from promega corporation madison, wi. Here, we present a rapid and sensitive assay based on the classical dualluciferase reporter technique which. Consecutive detection of firefly and renilla luciferase activities in a single sample.
In the dlr assay, the activities of firefly and renilla luciferases are measured sequentially from a single sample. Fast, sensitive detection of renilla luciferase activity. The dualluciferase reporter dlr assay system provides an efficient means of performing dualreporter assays. The components of the dlr assay system are identical in formulation to those provided with the dlr assay system cat.
Using the synergyht multidetection microplate reader to. Promega dualluciferase reporter dlr assay systems dual. This kit is a new and improved replacement for the firefly luciferase assay kit catalog no. Any search on dualluciferase reporters usually generates a lot of returns.
Such systems have found widespread application through the availability of commercial assay kits the dualluciferase reporter assay system, cat. Dualluciferase reporter assay and dualluciferase reporter assay system quick protocol fb024 pdf 73. Dualluciferase reporter assay system harvard university. Dualluciferase reporter assay and dualluciferase reporter assay system quick protocol fb024 pdf 73 kb english. The thermo scientific pierce luciferase dual assays deliver high sensitivity in a simple onestep detection protocol. 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 luciferase assay system was developed for reporter quantitation in mammalian cells. The dualluciferase reporter dlr assay system provides an efficient means of performing two reporter assays. Promega has several series of firefly and renilla luciferase vectors, pgl4, pgl3 and prl,designed for use with the dlr assay systems. 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. 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. So why are dualluciferase reporter assays so widely. Application of the dualluciferase reporter assay to the. Promega offers the pgl4 series of firefly and renilla luciferase vectors designed for.
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