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Dual-Light™ Luciferase & β-Galactosidase Reporter Gene Assay System (Invitrogen™)

The Dual-Light® luminescent reporter gene assay is designed for the rapid and sensitive detection of firefly luciferase and E. coli β-galactosidase in a single extract aliquot for gene expression assays.

• Detection of firefly luciferase and β-galactosidase in the same sample simplifies normalization of transfection efficiency.
• Wide dynamic range of β-galactosidase and luciferase assays enables accurate measurement of both enzymes from femtogram to nanogram range.
• Assay sensitivity is 100- to 1,000-fold better than either the isotopic⁄non-isotopic assays for chloramphenicol acetyl transferase (CAT) or the colorimetric⁄fluorescent assays for β-galactosidase, providing greater sensitivity than competing assay technologies.
• Highly sensitive assay with a wide dynamic range permits detection of high and low levels of reporter without performing numerous sample dilutions.
• Non-radioactive reporter gene assay kit eliminates concerns over use of radioisotopes.
• Assay can be completed in about one hour, providing fast assay turnaround.

Easy-to-Use
Light signal from each enzymatic reaction is measured sequentially in a luminometer with automatic injectors or other instrumentation in which light emission measurements can be performed within a short period. First, luciferase reporter enzyme activity is quantitated with an enhanced luciferase reaction. Following a 30-60 minute incubation and addition of a light-emission accelerator, β-galactosidase reporter enzyme activity is determined with Galacton-Plus® substrate. Both assays are combined into one simple assay using only one extract aliquot for greater convenience and precision. The entire assay is completed in less than one hour.

Wide Dynamic Range
The wide dynamic range of this dual assay enables accurate measurement of luciferase and β-galactosidase concentrations over seven orders of magnitude, from the femtogram to nanogram range. Colorimetric and isotopic reporter gene assays cannot rival the dynamic range of the Dual-Light® system. We have formatted this assay for tube or microplate luminometer, with injection capability.

Applications
Dual-Light reporter gene assay system has been very widely used for reporter quantitation⁄transfection normalization from transiently transfected mammalian cell lines as well as transfected primary cells. In addition, it has been used with a modified lysis buffer to quantitate luciferase and β-galactosidase activities from a novel reporter fusion construct in yeast cells. For these and additional references, please see our bibliography in the Literature⁄Resources section.

For Research Use Only. Not for use in diagnostics procedures.

Pierce™ Firefly Luc One-Step Glow Assay Kit (Thermo Scientific™)

The Thermo Scientific Pierce Firefly One-Step Luc Glow Assay Kit measures firefly luciferase activity in mammalian cell culture using a single reagent-addition step, making it ideal for high-throughput screening (HTS) applications.

Features of the Firefly One-Step Luc Glow Assay Kit:

Sensitive—highly sensitive detection of firefly luciferase activity
Odorless—reagents have no foul or toxic odor, eliminating the need to work in a fume hood
Convenient—one-step, mix-and-read protocol; no additional reagents required
Stable—increased signal half-life and stability compared to flash assays
Simple—does not require a luminometer with injectors
Automation-friendly—amenable to high throughput assays

This glow-type luciferase assay kit is designed for use with cultured mammalian cells that are transfected with vectors expressing the appropriate firefly luciferase reporter enzyme. Several features make the kit especially suitable for luminometers without injectors and for high-throughput screening (HTS) applications. The convenient, one-step, homogenous protocol minimizes handling steps to support the use of automation. Simply combine the supplied substrate solution and assay buffer to make a single working solution, and then add it to microplate wells containing the transfected, treated cells. The stable signal output (glow) of this system provides flexibility with regard to incubation time. When used with Thermo Scientific Firefly Luc Plasmids, the kit provides a highly sensitive bioluminescent reporter assay system for the detection of promoter or pathway activity.

Includes:
Assay buffer and substrate solution (100X)

Requires:
Firefly luciferase and luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers.

Applications:
• Promoter studies for analyzing cis- and trans-acting regulatory elements
• Drug screening
• siRNA and miRNA screening
• Multiplexed assays to study off-target effects
• Protein localization reporter assays
• Signal transduction pathway analysis

Firefly luciferase is a 62kDa protein produced in nature by several species of the Lampyridae family of beetles, which includes the genera Photinus and Luciola. Thermo Scientific Red Firefly Luciferase has an emission range of 560-700nm (λmax = 613nm). Bioluminescent signal from firefly luciferase originates from the oxidation of D-Luciferin. Light output captured using a luminometer can be correlated with the amount of firefly luciferase protein produced and used to determine the activity of the promoter driving firefly luciferase expression. Firefly luciferase bioluminescence provides researchers with a non-radioactive assay for measuring transcriptional activity of regulatory elements. Firefly luciferase reporter activity can be used to determine promoter activity, to study upstream pathway activity in the cell, or to determine effect of activators or inhibitors.

FITC-Casein for Pierce™ Fluorescent Protease Assay Kit (Thermo Scientific™)

FTC-Casein, 2.5 mg, lyophilized, FRET/FP Certified Grade, for use with the Pierce Fluorescent Quantitative Protease Assay Kit.

Related Products
Pierce™ Colorimetric Protease Assay Kit
Pierce™ Fluorescent Protease Assay Kit

RediPlate™ 96 EnzChek™ Tyrosine Phosphatase Assay Kit (Invitrogen™)

The RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit is a ready-to-use, fluorescence-based microplate assay for detecting tyrosine phosphatases (PTPase) and their corresponding modulators and inhibitors. Each RediPlate 96-well microplate has removable lanes, enabling researchers to perform only as many assays as needed for each experiment or to perform high-throughput analysis.

Galacto-Light Plus™ β-Galactosidase Reporter Gene Assay System (Invitrogen™)

The Galacto-Light Plus™ Beta-Galactosidase Reporter Gene Assay System is designed for the rapid, ultrasensitive detection of β-galactosidase in reporter gene assays.

• The Galacto-Light Plus™ Beta-Galactosidase Reporter Gene Assay System is designed for luminometers without injectors.
• Wide dynamic range of β-galactosidase assay lets the user measure enzyme level accurately from femtogram to nanogram range.
• Assay sensitivity is 100- to 1,000-fold better than with either the isotopic⁄non-isotopic assays for chloramphenicol acetyl transferase (CAT) or the colorimetric⁄fluorescent assays for β-galactosidase, providing greater sensitivity than competing assays.
• Highly sensitive assay with a wide dynamic range permits detection of high and low levels of reporter without performing numerous sample dilutions.
• Non-radioactive reporter gene assay kit eliminates concerns over use of radioisotopes.
• Assay can be completed in about one hour, providing fast assay turnaround.

Sensitive
This chemiluminescent assay exhibits over three orders of magnitude greater sensitivity compared to colorimetric β-galactosidase assays and can be completed in a fraction of the time required for assays for chloramphenicol acetyltransferase (CAT) and ELISA-based reporter assays.

Ready-to-Use
The Galacto-Light™ Plus™ system incorporates chemiluminescent substrates, reaction buffer, and light-emission accelerator containing luminescence enhancers, in a ready-to-use format. The reaction buffer is supplied at a pH that aids in discrimination of endogenous β-galactosidase, while having little effect on the bacterial form of the enzyme. This enables sensitive detection even in cell lines with relatively high levels of endogenous β-galactosidase activity.

Products described herein are guaranteed for 12 months from the date of purchase.

For Research Use Only. Not for use in diagnostics procedures.

NA-XTD™ Influenza Neuraminidase Assay Reagent Set, no plates included (Invitrogen™)

The NA-XTD™ Influenza Neuraminidase Assay Kit is a next-generation chemiluminescence-based assay that provides a longer signal read-out compared to the first-generation NA-Star® Influenza Neuraminidase Inhibitor Resistance Detection Kit. The NA-XTD™ kit includes detection reagents and microplates, eliminating the need for luminometers equipped with a reagent injector, and improving ease-of-use. This kit also includes complete assay protocols for quantitating sensitivity to neuraminidase inhibitors in various influenza virus isolates including: human influenza types A and B, A⁄H1N1 pandemic, avian, equine and porcine viruses.

Key product features:
• Superior performance—improved, long-lived light emission kinetics and read-time flexibility
• High sensitivity—high assay signal-to-noise, detection of low-virus concentrations, and wide assay dynamic range
• Simple instrumentation requirement—no reagent injectors or luminometer needed
• Multiple applications—neuraminidase inhibitor IC50 assays and cell-based virus quantitation

Long-Lived Light Emission Kinetics & Read-Time Flexibility
The NA-XTD™ assay yields much longer-lived light emission kinetics than does the NA-Star® assay, eliminating the need for luminometer instrumentation with reagent injectors and enabling read-time flexibility and batch-mode processing of assay plates. NA-XTD™ assay signal half-life is approximately 2 hours, longer than the 5-10 minute half-life of the NA-Star® assay signal. IC-50 values determined from data collected immediately or up to 3 hours following addition of NA-XTD™ Accelerator solution are identical (Figure 1).

High Sensitivity
The NA-XTD™ chemiluminescent assay provides higher detection sensitivity (better low-end detection limit), higher assay signal-to-noise ratio, and wider assay dynamic range than fluorescent assays with the MUNANA substrate. The NA-XTD™ assay also typically demonstrates slightly higher signal-to-noise than the NA-Star® assay. The NA-XTD™ assay provides 2–50-fold higher sensitivity by signal-to-noise ratio than MUNANA-based fluorescence assays, depending on the virus isolate (Figure 2). The NA-XTD™ assay provides a dynamic range of detection of 3–4 orders of magnitude of neuraminidase enzyme concentration, compared to 2–3 orders of magnitude range with fluorescent MUNANA assays. The wide chemiluminescent neuraminidase assay range enables determination of IC-50 values over a wide range of virus concentrations, eliminating the need to titer virus prior to performing IC50 determination assays (Figure 3).

Simple Instrumentation Requirement
The NA-XTD ™ assay can be used for virus quantitation in media samples from 96-well microplates or other virus cultures for monitoring viral growth or infection, or for performing viral inhibition assays in a cell-based system. Optimally, a small sample of culture media is removed and assayed directly with NA-XTD™ reagents, permitting multiple samples to be assayed over time (Figure 4).

Multiple Applications in One Complete Kit
The NA-XTD™ Assay Buffer is used as diluent for virus samples, neuraminidase inhibitors and NA-XTD™ substrate. An optional NA Sample Prep Buffer is also included for Triton® X-100 detergent addition to virus preparations, which increases NA activity in some virus preparations. The next-generation NA-XTD™ Accelerator solution triggers high intensity light emission from the NA-XTD™ reaction product. For added convenience and assay performance, the kit includes NA-Star™ Detection Microplates. These 96-well solid white assay microplates were selected for optimum assay performance, including high signal intensity, low background, and minimum well-to-well cross-talk. The kit includes a comprehensive assay protocol that provides virus and neuraminidase inhibitor (NI) dilution recommendations, a recommended plate layout for NI sensitivity assays, a protocol for virus quantitation, and a literature reference list. The NA-XTD™ Influenza Neuraminidase Assay Kit is compatible with a wide range of luminometers, including single-mode and multi-mode instruments, with no need for injectors.

For Research Use Only. Not for use in diagnostics procedures.

Z'-LYTE™ Kinase Assay Kit - Ser/Thr 22 Peptide

A coumarin and fluorescein-labeled peptide substrate which can be used for kinase screening in an antibody-free format. This peptide is based upon CDK7, containing Ser-161.

Yeast beta-Galactosidase Assay Kit (Thermo Scientific™)

Thermo Scientific Yeast β-Galactosidase Assay Kit is specially formulated for yeast cells and ideal for identifying protein interactions in-vivo using two-hybrid systems.

Features of Yeast beta-Galactosidase Assay Kit:

• Efficient lysis of yeast cells and a colorimetric detection system
• Quantitative or qualitative assay for yeast colonies grown on solid media or in suspension
• Colorimetric assay easily measured in a plate reader or spectrophotometer (420nm)
• Adaptable to 96-well microplates or tube assay formats
• Allows cells obtained from solid media or liquid culture to be assayed directly with no harvesting or washing steps.• Perform homogeneous screening assays or measure activity secondarily after cell lysis
• Also compatible with prepared lysates from other species such as bacteria

Yeast β-Galactosidase Assay Kit allows qualitative or quantitative determination of β-galactosidase activity in solution directly from colonies growing on solid medium. A portion of the colony is suspended in a mixture of Thermo Scientific Y-PER Yeast Protein Extraction Reagent and β-Galactosidase Assay Buffer. After a brief incubation period, the solution turns yellow from the hydrolysis of o-nitrophenyl-β-D-galactopyranoside (ONPG) to o-nitrophenol (ONP) and galactose in a mildly alkaline solution. The assay becomes quantitative if calibrated to cell number by measuring the optical density at 660nm.

The gene encoding β-galactosidase (lacZ) of E. coli has been widely used as a reporter gene in many different prokaryotic and eukaryotic organisms. β-Galactosidase activity is also commonly used as an indicator of protein-protein interactions in-vivo using two-hybrid systems. The interaction strength is verified or quantitated using a β-galactosidase activity assay.

Z'-LYTE™ Kinase Assay Kit - Ser/Thr 17 Peptide

A coumarin and fluorescein-labeled peptide substrate which can be used for kinase screening in an antibody-free format. This peptide is based upon glycogen synthase, containing Ser-7.

Pierce™ Renilla-Firefly Luciferase Dual Assay Kit (Thermo Scientific™)

The Thermo Scientific Pierce Renilla-Firefly Luciferase Dual Assay Kit provides the necessary reagents to simultaneously detect intracellular Renilla and Red Firefly luciferase activity in mammalian whole cell lysates.

Features of the Renilla-Firefly Luciferase Dual Assay Kit:

Simultaneous—concurrent, filter-based, wavelength-separated detection of two luciferase activities
Sensitive—measure Green Renilla and red firefly luciferase activities in the same sample
Fast—no quenching step required, unlike in traditional sequential dual assays
Multiplex—capable of quantitating two cellular activities in the same sample(s)

The Pierce Renilla-Firefly Luciferase Dual Assay Kit is a highly sensitive assay that allows for the simultaneous detection of Green Renilla and Red Firefly luciferase activity. Green Renilla luciferase acts as an experimental reporter with constitutively active red firefly as a normalization control. As shown in the animation video below, this reporter-and-control combination enables simultaneous monitoring of experimental reporter and control luciferase activities in a single-read assay without the need for two-step addition of substrate reagents or quenching. The assay working solution contains substrates for both luciferases, and the reactions occur simultaneously with flash-type kinetics. The resulting luminescent signals are spectrally resolvable using filters. In a single sample, researchers can assay transcriptional activity of regulatory elements, signal transduction pathways, and effects of activators or inhibitors.

Includes:
Cell lysis buffer, reaction buffer, and substrates coelenterazine and D-luciferin

Requires:
Green Renilla and Red Firefly luciferase reporters; Filter sets at 525nm±20nm for Green Renilla Luc and 615 to 675nm for Red Firefly Luc. Luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers; Injector required for assessing more than 24 wells at a time.

Applications:
• Study two regulatory elements at the same time
• Monitor two signaling pathways simultaneously
• Enable studying more than one target per screen (e.g. off-target effects)

Green Renilla produces brighter bioluminescent signal than firefly and native Renilla luciferases. The bioluminescent signal (λmax= 535nm) produced by Green Renilla luciferase protein results from the oxidation of coelenterazine. The second assay uses red firefly luciferase, which is a mutant form of the Japanese firefly luciferase Luciola cruciata. This luciferase produces a red-shifted emission spectrum (λmax= 613nm) that results from the oxidation of D-luciferin in the presence of ATP.

More Product Data
Simultaneous dual-emission detection of luciferase reporter assays

Related Products
Pierce™ Renilla-Firefly Luciferase Dual Assay Kit

Z'-LYTE™ Kinase Assay Kit - Ser/Thr 20 Peptide

A coumarin and fluorescein-labeled peptide substrate which can be used for kinase screening in an antibody-free format. This peptide is based upon a synthetic peptide substrate for NDR1.

Pierce™ Colorimetric Protease Assay Kit (Thermo Scientific™)

The Thermo Scientific Pierce Colorimetric Protease Assay Kit measures total protease activity in samples, providing a means to assess the progress of protease isolation procedures or quantify protease contamination in biological samples. This kit uses fully succinylated casein as a substrate for the assay. Hydrolysis of this substrate in the presence of protease results in the release of peptide fragments with free amino-terminal groups. These peptides are reacted with trinitrobenzene sulfonic acid (TNBSA), followed by measurement of the absorbance increase that results from the formation of yellow colored TNB-peptide adducts.

Features of the Colorimetric Protease Assay Kit:

Casein substrate—measure activity of any protease that cleaves casein into peptide fragments
Trypsin standard—quantify protease activity relative to trypsin, a universally accepted reference
Sensitive—1000 times more sensitive than assays that use unmodified forms of casein
Simple and fast—test tube and microplate protocols are completed in less than one hour
Homogenous—addition steps only; no separation, transfer or stop steps required
Customizable—easily adapt time, temperature and pH to optimize sensitivity for proteases of interest

Applications:
• Assess functional integrity and overall progress of protease purification
• Quantify protease contamination in protein samples
• Characterize the activity of unusual proteases compared to trypsin
• Evaluate buffer conditions for their affects on protease function and activity

Additional Specifications:
• Substrate: succinylated casein (not available separately)
• Basis of assay: detection of primary amines resulting from proteolysis
• Detection reagent: trinitrobenzene sulfonic acid (TNBSA)
• Measurement: absorbance at 450nm (plate reader or spectrophotometer)

Related Products
Pierce™ Fluorescent Protease Assay Kit
FITC-Casein for Pierce™ Fluorescent Protease Assay Kit

Z'-LYTE™ Kinase Assay Kit - Ser/Thr 4 Peptide

A coumarin and fluorescein-labeled peptide substrate which can be used for kinase screening in an antibody-free format. This peptide is based upon glycogen synthase, containing Ser-7 and a consensus seq Hyd-X-R-X-X-S⁄T-X-X, where Hyd is bulky hydrophobic, F > L > V>>A, and X is any residue.

Gal-Screen™ β-Galactosidase Reporter Gene Assay System for Mammalian Cells (Invitrogen™)

The Gal-Screen® assay system combines direct cell lysis with rapid, ultra-sensitive chemiluminescent detection of β-galactosidase reporter enzyme.

• Homogeneous assay format allows detection of β-galactosidase in the presence of normal culture media without removal of media and without an additional cell lysis step, providing the easiest and most streamlined assay procedure possible.
• The kinetics of the glow-assay provides a "window" during which measurements may be performed, facilitating HTS applications where assay automation is used.
• Can be used with either mammalian or yeast model systems, providing flexibility in choice of model systems.
• Wide dynamic range of β-galactosidase assay enables accurate measurement of enzyme from femtogram to nanogram range.
• Assay sensitivity is 100 to 1,000-fold better than either the isotopic⁄non-isotopic assays for chloramphenicol acetyl transferase (CAT) or the colorimetric⁄fluorescent assays for β-galactosidase, providing greater sensitivity than competing assay technologies.
• Highly sensitive assay with a wide dynamic range permits detection of high and low levels of reporter without performing numerous sample dilutions.
• Non-radioactive reporter gene assay kit eliminates concerns over use of radioisotopes.
• Assay can be completed in about one hour, providing fast assay turnaround.

Ideal for Screening
This homogeneous assay is ideally suited for screening applications where assay automation is required. The Gal-Screen® system uses Galacton-Star® chemiluminescent substrate for convenient measurement in a luminometer. Light emission reaches maximum in 60-90 minutes and remains constant for 45-90 minutes.

For Research Use Only. Not for use in diagnostics procedures.

Pierce™ Renilla Luciferase Glow Assay Kit (Thermo Scientific™)

The Thermo Scientific Pierce Renilla Luciferase Glow Assay Kit provides an extremely bright and stable bioluminescence signal (half-life approx. 3 hours), especially in the presence of Green Renilla luciferase reporter.

Features of the Renilla Luciferase Glow Assay Kit:

Sensitive—highly sensitive detection of Green Renilla luciferase activity
Stable—increased signal stability compared to flash assays
Simple—does not require a luminometer with injectors
Compatible—assay reagents compatible with other Renilla luciferases
Automation-friendly—amenable to high throughput assays
Convenient—contains a universal cell lysis buffer and optimized glow assay reagent
Safe—allows one to perform non-radioactive assays

This Pierce Luciferase Glow Assay Kit contains reagents for measuring the activity of Renilla luciferase in cell lysates. In the presence of luciferase, the glow reagent provides stable bioluminescent signal and is highly suited for luminometers without injectors or for batch processing of samples. The light output generated by the luciferase reaction can be correlated with the amount of luciferase protein produced which in turn is proportional to the promoter activity driving the luciferase expression. The glow assay reagents were optimized for best results with Thermo Scientific Green Renilla Luc Plasmids; however, this glow kit may be used to detect the activity of other Renilla luciferase derivatives that use coelenterazine as a substrate.

Includes:
Cell lysis buffer, reaction buffer and substrate

Requires:
Renilla luciferase and luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers.

Applications:
• Promoter studies for analyzing cis- regulatory elements and trans-acting factors
• Drug screening
• siRNA and miRNA screening
• Multiplexed assays to study off-target effects
• Protein localization reporter assays
• Signal transduction pathway analysis
• RNA splicing studies

The Green Renilla luciferase is a 36kDa protein produced by a derivative of the wild type Renilla luciferase gene from the sea pansy, Renilla reniformis. Compared to the wild type luciferase, Green Renilla is more stable in serum and has an the emission spectrum that is shifted toward the green region. The protein provides extremely bright flash signal that decays rapidly. The Pierce Renilla Glow Assay Kit reagent effectively stabilizes this bright bioluminescence to yield a signal half-life greater than 3 hours, thus eliminating the need for a luminometer with injectors. The assay kit was optimized for best results with Thermo Scientific Green Renilla Luc Plasmids; however, this glow kit may be used to detect the activity of other Renilla luciferase derivatives that use coelenterazine as a substrate.

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Pierce™ Renilla-Firefly Luciferase Dual Assay Kit