Navigation Links
Dual-Luciferase Quantitation in the LMax Microplate Luminometer (MaxLine Application Note #39)

Evelyn McGown, Ph.D. and Michael Su, M.S.
Molecular Devices Corporation, 8/00


INTRODUCTION
Reporter gene assays are used to study eukaryotic gene expression. Dual geneticreporters are commonly used in transient transfections of cultured cells tominimize experimental variability caused by differences in cell number, viabilityor transfection efficiency. One vector containing the experimental reporter gene iscotransfected with a second vector containing a different reporter gene to serve asthe control. Typically, the experimental reporter is coupled to a regulatedpromoter, and the control reporter gene is coupled to a constitutive promoter. Bynormalizing the changes in expression of reporter activity to the activity of thecontrol, experimental variability can be minimized.

Colorimetric reporter assays have been widely used, but bioluminescent reportersystems, especially using firefly luciferase, are becoming increasingly popularbecause of their speed, sensitivity and wide dynamic range. PromegaCorporation has recently introduced their Dual-Luciferase Reporter Assay System which utilizes the firefly (Photinus pyralis) and Renilla (Renilla reniformis)luciferases1. Because the enzymes have different substrate requirements, they canboth be measured in a single reaction mixture. The firefly luciferase is typicallyused as the experimental reporter, and the renilla luciferase serves as the control.

Figure 1 illustrates the two enzymatic reactions. Firefly luciferase enzymecatalyzes the oxidation of luciferin with the concomitant release of light2. The reaction requires ATP, Mg+2 and O2. Renilla luciferase catalyzes the O2-dependentoxidation of coelenterate luciferin (coelenterazine) but does not require Mg orATP3. Both reactions can easily be measured in the Molecular Devices Lmaxmicroplate luminometer.
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. Quantitation of Cabergoline at Extremely Low Plasma Concentrations with a Triple Quadrupole Mass Spectrometer
2. Bioanalytical Method Intraday Validation for the Quantitation of Paroxetine in Bovine Plasma using the TSQ Quantum Mass Spectrometer
3. Quantitation of the Protein MIA as a Marker for Chondrocytes in Research Samples
4. Comparison of Different Methods of mRNA Quantitation
5. Quantitation of DNA for Automated Sequencing Using the VersaFluor Fluorometer
6. Quantitation of RNA for Hybridization Blots Using the VersaFluor Fluorometer
7. Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
8. Quantitation of GAD67 Gene Expression in Prefrontal Cortex of Schizophrenia Patients Using the iCycler iQ Detection System and Molecular Beacons, Rev A
9. Benchmark Plus Microplate Reader: Quantitation of Protein Concentration Using Two Different Colorimetric Assay Kits, Rev A
10. A Robust LC/MS/MS Method for the Quantitation of Very Long Chain Fatty Acids for Research into the Characterization of Peroxisomal Disorders
11. A new LC/MS/MS Research Method for Rapid Quantitation of Five Immunosuppressant Drugs, including Mycophenolic Acid (MPA)
Post Your Comments:
(Date:2/26/2015)... , Feb. 26, 2015   Regulus Therapeutics ... company leading the discovery and development of innovative medicines ... , Ph.D., President and Chief Executive Officer of Regulus, ... Company 35 th Annual Healthcare Conference on Wednesday, ... is being held at the Boston Marriott Copley Place.  ...
(Date:2/26/2015)... CA (PRWEB) February 26, 2015 The ... online demonstration to support life extension technologies and awareness. ... UTC, Universal Coordinated Time, on March 21st 2015, in ... world. , Eric Schulke, Founder of MILE, says, ... act with urgency. The Movement for Indefinite Life Extension ...
(Date:2/26/2015)... -- The healthcare landscape is rapidly and radically changing. To ... host its 20 th Anniversary Medical Technologies 2015: ... 10, at the Hilton San Diego Resort and Spa in ... event is a platform for global medical devices, diagnostics, life ... how to succeed in an uncertain future. Participants ...
(Date:2/26/2015)... February 26, 2015 Already well respected ... line by expanding its Length Gauge offerings with new ... , Specifically, HEIDENHAIN’s pencil probe-style SPECTO length ... specification. Two new 1Vpp, 12mm stroke length variants ... applications. One is a variant with a weak spring, ...
Breaking Biology Technology:Regulus to Present at Cowen and Company 35th Annual Healthcare Conference 2Regulus to Present at Cowen and Company 35th Annual Healthcare Conference 3Indefinite Life Extension Activists Organize Online Demonstration 2Indefinite Life Extension Activists Organize Online Demonstration 3Embrace the Future of Medical Technologies with Frost & Sullivan 2Embrace the Future of Medical Technologies with Frost & Sullivan 3Accuracy and Versatile Gauging Go Hand in Hand at HEIDENHAIN 2
... Quest PharmaTech Inc.,("Quest" or the "Company") is ... support from NRC-IRAP towards the development of ... Sonodynamic Therapy arises out of the Company,s ... and skin diseases. The,Company,s lead compound, SL017, ...
... LLC funds Company Initial $250,000 under March ... ... bioMETRX Inc., (OTC,Bulletin Board: BMRX), a leading developer and supplier of ... the assets and former personnel of Sequiam,Corporation (OTC Bulletin Board: SQUME) ...
... HAVEN, Conn., April 22 Rib-X Pharmaceuticals, ... focused on the,discovery and development of novel ... announced that Graham Johnson,Ph.D., Chief Research Officer ... the Cambridge Healthtech Institute,s 2nd Annual "The,Challenge ...
Cached Biology Technology:Quest PharmaTech Receives Support from National Research Council's Industrial Research Assistance Program (NRC-IRAP) to develop Ultrasound Technology for the Treatment of Cancer 2bioMETRX, Inc. Initiates Integration of Personnel and Assets of Sequiam Corp. 2Rib-X Pharmaceuticals to Present at Cambridge Healthtech Institute's 2nd Annual Challenge of Antibacterial Development Conference 2
(Date:2/10/2015)...  Alere Inc. (NYSE: ALR), a global leader ... the quarter ended December 31, 2014. ... Alere said, "We made substantial progress in the ... leader in rapid diagnostics and delivering against our ... in early January enabled us to substantially reduce ...
(Date:2/5/2015)... RESEARCH TRIANGLE PARK, North Carolina , ... in sein 35. Jahr als spezialisiertes Logistikunternehmen ... seine Position als einzige klinische Logistikfirma (Clinical ... soll. Die Kernbotschaft der neuen Kampagne lautet ... Protokollen und Lieferungen. Foto: ...
(Date:2/5/2015)... , Jan. 26, 2015   Epic Sciences , ... cancer, today announced that Murali Prahalad , Ph.D., president ... Medicine World Conference (PMWC) 2015: Silicon Valley, which is ... Mountain View, Calif. on January 26-28, 2015. ... Come of Age as Biomarkers." Last year, Epic Sciences was ...
Breaking Biology News(10 mins):Alere Inc. Announces Fourth Quarter 2014 Results 2Alere Inc. Announces Fourth Quarter 2014 Results 3Alere Inc. Announces Fourth Quarter 2014 Results 4Alere Inc. Announces Fourth Quarter 2014 Results 5Alere Inc. Announces Fourth Quarter 2014 Results 6Alere Inc. Announces Fourth Quarter 2014 Results 7Alere Inc. Announces Fourth Quarter 2014 Results 8Alere Inc. Announces Fourth Quarter 2014 Results 9Alere Inc. Announces Fourth Quarter 2014 Results 10Alere Inc. Announces Fourth Quarter 2014 Results 11Alere Inc. Announces Fourth Quarter 2014 Results 12Alere Inc. Announces Fourth Quarter 2014 Results 13Alere Inc. Announces Fourth Quarter 2014 Results 14Alere Inc. Announces Fourth Quarter 2014 Results 15Alere Inc. Announces Fourth Quarter 2014 Results 16Alere Inc. Announces Fourth Quarter 2014 Results 17Alere Inc. Announces Fourth Quarter 2014 Results 18Alere Inc. Announces Fourth Quarter 2014 Results 19Alere Inc. Announces Fourth Quarter 2014 Results 20Alere Inc. Announces Fourth Quarter 2014 Results 21Alere Inc. Announces Fourth Quarter 2014 Results 22Alere Inc. Announces Fourth Quarter 2014 Results 23Alere Inc. Announces Fourth Quarter 2014 Results 24Alere Inc. Announces Fourth Quarter 2014 Results 25Alere Inc. Announces Fourth Quarter 2014 Results 26Alere Inc. Announces Fourth Quarter 2014 Results 27Alere Inc. Announces Fourth Quarter 2014 Results 28Alere Inc. Announces Fourth Quarter 2014 Results 29Alere Inc. Announces Fourth Quarter 2014 Results 30Alere Inc. Announces Fourth Quarter 2014 Results 31Alere Inc. Announces Fourth Quarter 2014 Results 32Alere Inc. Announces Fourth Quarter 2014 Results 33Alere Inc. Announces Fourth Quarter 2014 Results 34Alere Inc. Announces Fourth Quarter 2014 Results 35Alere Inc. Announces Fourth Quarter 2014 Results 36
... has been found on the surface of a type of ... and purify adult stem cells for use in therapies aimed ... UT Southwestern Medical Center report. , The molecule, called SSEA-4, ... bone marrow that give rise to fat, cartilage and bone. ...
... scientists has published the first two genome sequences ... Phytophthora--a name that literally means "plant destroyer." The ... "fy-TOFF-thor-uh") attack a broad range of plants and ... hundreds of billions of dollars each year. , ...
... Diego have discovered that the presence of a key protein ... at center due to a fluorescent protein) allows the formation ... unit eyes, an evolutionary change to an "open system" that ... angular sensitivity. In contrast, beetles (shown surrounding the fruit fly), ...
Cached Biology News:Molecular 'marker' on stem cells aids research, perhaps therapies 2Genome info from 'plant destroyers' could save trees, beans and chocolate 2
Goat polyclonal to PINX1 ( Abpromise for all tested applications). entrezGeneID: 54984 SwissProtID: Q548A5...
... membrane-filtered isotonic and enzyme-free aqueous formulations ... agents in either Ca2+- and Mg2+-free ... Ca2+- and Mg2+-free phosphate-buffered saline for ... support substrates and each other for ...
...
... , 2 x 50 ml 10X TBST Wash Buffer ... ml BL21 glycerol stock , 0.2 ml BLT5403 glycerol ... 1010 pfu T7Select Positive Control Lysate , 1010 pfu ... Conjugate , 2 ml T7Select S-protein , 50 ...
Biology Products: