Shop All Enzyme Detection & Activity Assays

EnzChek™ Phospholipase A2 Assay Kit (Invitrogen™)

This kits takes our stand alone assay for phospholipase A2, PLA2 (A10072) and combines the necessary Ez and lipids to run 2 complete 96 well microplate assays that monitor activity in purified enzyme preparations and cell lysates. The EnzChek Phospholipase A2 Kit provides enough reagents for 2 microplates, using 200 µl volumes in 96 well format to perform continuous fluorometric monitoring of PLA2 . This product offers an alternative to our (B7701), (bis-BODIPY® FL C11-PC) reagent, by providing an PLA2 selective substrate and one that is ratiometric, thereby lowering variations produced by instrumentation and assay conditions. Phospholipase A2 or PLA2 represents a family of enzymes that hydrolyze the sn-2 ester linkage of phospholipids. The activities of these enzymes play important roles in cardiovascular, inflammatory and nervous system disorders, and in cancers. The EnzChek® Phospholipase A2 substrate provides sensitive and continuous rapid real-time monitoring of PLA2 enzyme activities. This unique substrate is selective for PLA2 and can be used either in a intensity or ratiometric based detection mode. In intensity based detection mode the PLA2 activity is monitored by the intensity increase of a single wavelength at approximately 515 nm. In ratiometric analysis, which is based on the distinct fluorescence resonance energy transfer (FRET) emission of this substrate prior to and after cleavage, PLA2 is detected by changes in the emission intensity ratio at 515/575 nm with excitation at ≈ 460 nm. Either detection mode provides a simple method with low background, high sensitivity and high specificity for PLA2.

LanthaScreen™ TR-FRET BACE1 Assay Kit

BACE1 (beta-secretase) is a key enzyme involved in the production of amyloid beta-peptides found in extracellular amyloid plaques of Alzheimer’s disease (AD). In some cases, early-onset familial AD can be attributed to a "Swedish"mutation in the amyloid precursor protein (APP), which dramatically enhances the cleavage of this protein by BACE1. This and other genetic and pathological evidence has led to therapeutic approaches that focus on the inhibition of BACE1 and other APP-cleaving enzymes, such as gamma-secretase.

Invitrogen’s LanthaScreen® TR-FRET BACE1 assay provides sensitive high-throughput screening for potential inhibitors of beta-secretase. The kit uses a terbium (Tb)-labeled anti-biotin antibody and a fluorescein-labeled BACE1-biotin substrate in a homogeneous TR-FRET assay format (Figure 1).



Contents and Storage:

The LanthaScreen® TR-FRET BACE1 Assay Kit contains BACE1 protein, fluorescently labeled BACE1 substrate, Tb-labeled anti-biotin antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

EnzChek™ Protease Assay Kit, red fluorescence, 100-1000 assays (Invitrogen™)

The EnzChek® Protease Assay Kit, red fluorescence, is a fast, simple, and direct fluorescence-based assay for detecting metallo-, serine, acid, and sulfhydryl proteases. The accompanying increase in fluorescence, which can be measured with a fluorescence microplate reader, is proportional to protease activity.

See our complete line of Fluorescence Microplate assays.

• Generalized mechanism allows for measurement of a variety of proteases
• Can be used to continuously measure protease kinetics
• Excitation/emission similar to Texas Red® dye (589/617 nm)

This EnzChek® Protease Assay Kit contains a casein derivative that has been extensively labeled with the red-fluorescent BODIPY® TR-X dye, which results in a quenching of the fluorescent dye. Protease-catalyzed hydrolysis releases the highly-fluorescent BODIPY® TR-X dye-labeled peptides, allowing for quantitative detection of protease activity in solution. The red-fluorescent BODIPY® TR-X dye has excitation and emission spectra similar to those of the Texas Red® fluorophore.

EnzChek™ Phospholipase A1 Assay Kit (Invitrogen™)

This kits takes our stand-alone assay for phospholipase A1, PLA1 (A10070) and combines the necessary reagents to run 2 to 10 complete 96 well microplate assays that monitor activity in purified enzyme preparations and cell lysates. The importance of phospholipases in cellular signaling, lipid metabolism, inflammatory responses and pathological disorders related to these processes has stimulated demand for fluorescence-based activity monitoring methods. In particular the phospholipases resident in plasma and endothelium can perturb circulating LDL and HDL particles, creating pro-artherogenic forms. Recent evidence is drawing a link between these lipases and the progression of several severe neurodegenerative diseases, including Alzheimer's. Molecular Probes’ fluorogenic phospholipase A1 substrates is designed to provide continuous monitoring of phospholipase A1 (PLA1) in purified enzyme preparations, cell lysates and living cells. PLA-1 improves upon two existing reagents in two ways: first it is now PLA-1 specific and secondly it has improved the assay quality by decreasing initial background noise.

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.

Galacto-Star™ β-Galactosidase Reporter Gene Assay System for Mammalian Cells (Invitrogen™)

The Galacto-Star™ One-Step β-Galactosidase Reporter Gene Assay System chemiluminescent reporter gene assay system enables rapid, sensitive detection of β-galactosidase in mammalian cell lysates.

• Glow assay kinetics 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 lets the user measure enzyme level accurately 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 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
The chemiluminescent assay exhibits over three orders of magnitude greater sensitivity than 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.

Versatile
The Galacto-Star™ system includes Galacton-Star® chemiluminescent substrate for β-galactosidase and is suited for measurement in both luminometers and scintillation counters. Automatic injection is not required.

Long-Lasting Signal
Light emission from the Galacto-Star™ assay reaches maximum in 60-90 minutes and remains constant for nearly one hour. Cell lysate is mixed with reaction buffer containing Galacton-Star® substrate and Sapphire-II™ luminescence enhancer. After a 60-90 minute incubation at room temperature, the signal is measured in a luminometer or scintillation counter.

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

β-Gal Staining Kit (Invitrogen™)

The β-Gal Staining Kit allows you to determine the percentage of transfected cells expressing lacZ. lacZ is a bacterial gene often used as a reporter construct in eukaryotic transfection experiments. The gene product of lacZ, β-galactosidase, is resistant to proteolysis in cellular lysates and its activity is easily assayed. β-galactosidase catalyzes the hydrolysis of X-gal, producing a blue color that can be easily visualized under a microscope.

Kinase Tracer 314, 25 µM

In the LanthaScreen® Eu Kinase Binding Assay, binding of an Alexa Fluor® conjugate or “tracer" such as Kinase Tracer 314 to a kinase is detected by addition of a Eu-labeled anti-tag antibody. Binding of the tracer & antibody to a kinase results in a high degree of FRET, whereas displacement of the tracer with a kinase inhibitor results in a loss of FRET. Unlike many kinase activity assays, this assay is a simple mix and read assay, with no development steps. Learn More and See Which Kinases Have Been Validated Using This Assay

Pierce™ Firefly Luciferase Flash Assay Kit (Thermo Scientific™)

The Thermo Scientific Pierce Firefly Luciferase Flash Assay Kit provides reagents to measure Red firefly luciferase activity in whole cell producing a very strong flash signal which decays rapidly.

Features of the Firefly Luciferase Flash Assay Kit:

• Highly sensitive detection of firefly luciferase activity
• Use with Pierce Firefly Luciferase Signal Enhancer for best results (maximum signal)
• Working solution stable for up to 2 months at -20°C
• Assay reagents compatible with other firefly luciferases
• Amenable to automation
• Contains a universal cell lysis buffer and optimized flash assay reagent
• Safe non-radioactive assay

Thermo Scientific Pierce Firefly Luciferase Flash Assay Kit contains reagents to measure the activity of firefly luciferase in mammalian whole cell lysates. Containing a convenient lysis buffer, assay buffer and firefly substrate, this kit provides a complete reagent system for the intracellular detection of promoter or pathway activity. Light output resulting from the luciferase reaction is captured using a luminometer and can be correlated with the amount of firefly luciferase protein produced and used to determine the activity of the promoter driving firefly expression. These reagents are optimized to work with Thermo Scientific Firefly Luc Plasmids; however, the Flash Assay Kit may be used to detect the activity of other firefly or ATP-dependent luciferases utilizing D-luciferin as a substrate.

Includes:
Cell lysis buffer, reaction buffer and substrate; purchase Luciferase Signal Enhancer separately.

Requires:
• Firefly luciferase and luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers.
• For best results (maximum signal), use with Pierce Firefly Luciferase Signal Enhancer

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
• Secretory pathway/protein localization reporter assays
• Signal transduction pathway analysis
• RNA splicing studies

Firefly luciferase is a 60kDa protein produced in nature by several species of the Lampyridae family of beetles which includes the genera Photinus and Luciola. 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 expression.

More Product Data
Monitoring neuronal differentiation using multiplexed luciferase reporters
Activation of the antioxidant response pathway by pesticide chemicals
Versatile luciferases: microplate luminometers and flash luciferase assays

Pierce™ Kinase Enrichment Kit with ADP Probe (Thermo Scientific™)

This Thermo Scientific Pierce Kinase Enrichment Kit utilizes an ActivX™ ADP Probe to covalently label the active site of ATPases, including chaperones and metabolic enzymes, to enable their selective enrichment using a desthiobiotin tag.

ActivX ADP Probes feature an amine-reactive nucleotide analog and a desthiobiotin (biotin analog) tag that facilitates selective labeling of lysines in the kinase active site and then subsequent enrichment and recovery of labeled protein. These features allow identification and profiling of target enzyme classes across samples or assessment of the specificity and affinity of enzyme inhibitors.

Features of the Pierce Kinase Enrichment Kit:

Specific – label only the conserved active-site lysines of nucleotide-binding proteins
Flexible – use for in vitro labeling of ATPase enzymes derived from cells or tissues
Compatible – use with Western blot or mass spectrometry (MS) workflows

Applications:
• Profile small-molecule binding affinities and active-site inhibition in a dose-dependent manner
• Identify dozens to hundreds of inhibitor targets and off-targets from tissues, cells and subcellular proteomes
• Enrichment of enzymes based on function

Thermo Scientific ActivX Desthiobiotin-ADP is a nucleotide derivative that covalently modifies the active site of enzymes at conserved lysine residues in the nucleotide binding site. The structure of these probes consists of a modified biotin (desthiobiotin) attached to the nucleotide through a labile acyl-phosphate bond. Desthiobiotin is a biotin analog that binds less tightly to biotin-binding proteins resulting in binding that is easily reversed by biotin displacement, low pH or heat denaturation.

Desthiobiotin-ADP probes can be used to selectively enrich, identify and profile target enzyme classes or assess the specificity of enzyme inhibitors. Because many ATPases and other nucleotide-binding proteins bind nucleotides or inhibitors even when they are enzymatically inactive, the desthiobiotin probes allow profiling of both inactive and active enzymes in a complex sample. Preincubation of samples with small-molecule inhibitors that compete for active sites can be used to determine inhibitor binding affinity. Active-site nucleotide probes also can be used to identify inhibitor off-targets.

These products are subject to a limited use label license.

Related Products
Pierce™ Kinase Enrichment Kit with ATP Probe
ActivX™ Desthiobiotin-ATP Probe
ActivX™ Desthiobiotin-ADP Probe

Z'-LYTE™ Kinase Assay Kit - Tyrosine 1 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 an optimal peptide substrate for SRC.

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

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.

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

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

Features of the Cypridina-Firefly Luciferase Dual Assay Kit:

Simultaneous—concurrent, filter-based, wavelength-separated detection of two luciferase activities
Sensitive—measure bright blue Cypridina 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 Cypridina-Firefly Luciferase Dual Assay Kit is a highly sensitive assay that allows for the simultaneous detection of Cypridina and Red Firefly luciferase activity. Cypridina luciferase acts as an experimental reporter with constitutively active red firefly as a normalization control. As shown in the animated videos 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 Vargulin and D-luciferin

Requires:
Cypridina and Red Firefly luciferase reporters; Filter sets in the ranges of 425 to 525nm for Cypridina 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)

Application Videos:
Cypridina luciferase produces brighter bioluminescent signal than firefly and native Renilla luciferases. The bioluminescent signal (λmax=463nm) produced by Cypridina luciferase results from the oxidation of Vargulin. This reaction does not require adenosine triphosphate (ATP) or other cofactors. Red firefly luciferase is a mutant form of the Japanese firefly luciferase from 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. Red firefly and Cypridina luciferases are used in the dual assay because the signals are spectrally resolvable.

More Product Data
Highly sensitive multiplex luciferase reporter assays
Simultaneous dual-emission detection of luciferase reporter assays
Monitoring neuronal differentiation using multiplexed luciferase reporters
Activation of the antioxidant response pathway by pesticide chemicals

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.