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

PREPARATION OF REAGENTS
1. Luciferase assay reagent II (LAR II) was prepared by resuspending the lyophilized luciferase assay substrate (E151A) in 10 mL luciferase assay buffer II (E1955).

2. The Stop & Glo reagent was prepared by first reconstituting the dried Stop & Glo substrate with 200 L Stop & Glo Substrate Solvent (E463A). After gently vortexing for 10 sec, the entire reconstituted substrate was mixed with 10 mL Stop & Glo buffer (E197A).


PREPARATION OF THE LMAX
1. The solvent delivery systems of the Lmax were both cleaned by pumping 75% ethanol through them and allowing it to stand for one hour, followed by rinsing generously with deionized water (5 washes, 99 cycles each).

2. The P injector was filled with the LAR II reagent by pumping 2 mL through it. The M injector was similarly primed with Stop & Glo Reagent.


ASSAY PROCEDURE
The kit instructions were followed, except that the measurements were made on diluted recombinant enzymes instead of lysed cells.

1. In SOFTmax PRO, Dual Read mode was selected. Both the P and M injectors were set to deliver 100 L/well, followed by a 2-second delay and a 5-second read.

2. The diluted firefly+Renilla luciferase standards and buffer blank were pipetted (20 L each) into microplate wells in quadruplicate. The final enzyme content for each ranged from 2.0 to 2 x 106 fg/well.

3. The plate containing the luciferase standards was placed into the drawer of the Lmax and the injection/read cycles initiated.

During the assay, the P injector supplies the LAR II reagent to each well a minimum of 1.6 sec ahead of the time the read head moves into position over that well. The M injector supplies Stop & Glo'"/>

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:2/27/2015)... A paper published today, " Financing ... in Science Translational Medicine, demonstrates the ... the risk associated with investing in the treatment ... of funding for developing so-called "orphan" drugs. ... between scientists at the National Center for Advancing ...
(Date:2/27/2015)... 27, 2015 The fully automated ... UC) can sequentially analyze up to 48 samples ... high-sensitivity detection of targets by mass spectrometry. ... fulfill the measurement requirements of a wide range ... drug delivery and search for disease biomarkers, additives ...
(Date:2/27/2015)... February 27, 2015 FamilyFarms Group is ... Manager of State Line Farms, received the 2015 Tomorrow’s ... producer, under the age of 35, who has demonstrated ... an honor to win the Tomorrow's Top Producer Horizon ... won the Top Producer Award and learning from their ...
(Date:2/27/2015)... BOSTON , Feb. 27, 2015   PureTech ... identifying, inventing and commercializing disruptive products and technologies in ... Christopher Viehbacher , former CEO and Member of the ... Directors.  "It,s has been a pleasure ... very excited that we will be working together more ...
Breaking Biology Technology:New Financing Technique May Hold Key to Unlocking Funding for Rare Diseases 2New Financing Technique May Hold Key to Unlocking Funding for Rare Diseases 3New Financing Technique May Hold Key to Unlocking Funding for Rare Diseases 4Shimadzu Corporation Launches Nexera UC - The World’s Foremost Unified Chromatography System 2Shimadzu Corporation Launches Nexera UC - The World’s Foremost Unified Chromatography System 3Matt Sims Receives the 2015 Tomorrow’s Top Producer Horizon Award 2Christopher Viehbacher, former CEO of Sanofi, Joins PureTech's Board of Directors 2Christopher Viehbacher, former CEO of Sanofi, Joins PureTech's Board of Directors 3Christopher Viehbacher, former CEO of Sanofi, Joins PureTech's Board of Directors 4
... Convergence, the architecture for voice, video and integrated data ... word virtual. It is no longer a murky technology ... eliminating boarders, physical boundaries, brick and mortar, convergence allows ... cost, thus increasing productivity as well as the bottom ...
... take advantage of the limitless minutes now included in ... a device that can significantly improve the quality of ... ,"When you've been talking on a cell phone for ... gets," notes Zhenqiang "Jack" Ma, an assistant professor of ...
... MADISON - Ohio State University's new "living laboratory" ... media-presentation system developed by Madison-based Sonic Foundry, ... view instructors' presentations, the company announced Monday. , ... to provide Ohio State with data for making ...
Cached Biology Technology:WTN Visions: Convergence is changing business 2WTN Visions: Convergence is changing business 3UW-Madison technology to advance cell phones 2UW-Madison technology to advance cell phones 3Ohio State's educational tech lab to use Sonic Foundry system 2
(Date:1/22/2015)... , Jan. 13, 2015  Today, FreeWavz ( ... launched its crowdfunding campaign on Fundable, https://www.fundable.com/freewavz ... build of production capacity to meet customer demand. ... Photo - http://photos.prnewswire.com/prnh/20150113/168790 ...
(Date:1/22/2015)... , Jan. 15, 2015  BellBrook Labs, a leader ... announced the launch of a TR-FRET (time resolved ... Transcreener UDP Assay, a high throughput screening assay ... assay will allow for sensitive detection of hundreds ...
(Date:1/22/2015)... , Jan. 20, 2015 NXT-ID, Inc. (NASDAQ: NXTD ... commerce market, reports on the recent success of the Wocket™ smart wallet ... Wocket smart wallet was named as one of the "11 Hot ... "5 Best Products Launched At CES So Far" by Newseveryday.com and "The ...
Breaking Biology News(10 mins):FreeWavz Launches on Fundable to Drive Speed-to-Market in 2015 2BellBrook Labs Launches a High Throughput Screening Assay for Glycosyltransferases with TR-FRET Readout 2CES Response for NXT-ID's Wocket Smart Wallet Kicks Off 2015 2CES Response for NXT-ID's Wocket Smart Wallet Kicks Off 2015 3
... today are genetically modified (GM) to resist insect pests ... one of the most commonly grown GM crops in ... on the long-term health of soils. Although there are ... potentially reducing insecticide use, there may be unintended side ...
... of scientists from the University of Missouri created a ... disease, an inherited neuromuscular disease affecting approximately 150,000 people ... a group of progressive disorders that affects the peripheral ... connects the brain and spinal cord to targets such ...
... /PRNewswire-iReach/ -- Global Information Inc. is pleased ... in the medical devices markets, driven by improving standards ... Renal Dialysis Equipment Market to 2017 The growing ... population are driving rapid growth in the number of ...
Cached Biology News:Genetically modified corn affects its symbiotic relationship with non-target soil organisms 2Genetically modified corn affects its symbiotic relationship with non-target soil organisms 3New genetically engineered mice aid understanding of incurable neuromuscular disease 2Global Surgical Devices Market Growth Driven by Improving Standards of Living and Longer Life Expectancies, Research Shows 2Global Surgical Devices Market Growth Driven by Improving Standards of Living and Longer Life Expectancies, Research Shows 3Global Surgical Devices Market Growth Driven by Improving Standards of Living and Longer Life Expectancies, Research Shows 4
Human FGF R2 (alpha isoforms) MAb (Clone 98706)...
dithiothreitol, sulfhydryl reducing agent, C4H10O2S2; MW: 154.24, 97% pure (minimum), CAS No: 27565-41-9, store at -20 C ...
... Nucleon HT, 1 kit. Facilitates the simple, ... sections and hard tissue requiring proteinase K ... quality DNA from 50 preparations of up ... DNA/RNA Synthesis & Purification, Purification and Amplification ...
Human LT beta R/TNFRSF3 Phycoerythrin MAb (Clone 71319)...
Biology Products: