for luciferase reporter assays - Fisher Scientific · gene and then transferring the resulting...

pMCS- Luc

Spe IEco RI*Acc65IKpn ISac IXho IHind IIIBam HI

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pUC Ori

Puromycin R SV40 Ori/Promoter BGH pA

Luc

SV

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A

AmpR Ter

MCS

Lac01 TPS

pCMV-LucpUC Ori

Amp R

SV40 pA Puromyc

inR

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Ori

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PCMV Luc

BGH pA

Lac01 TPS

Stronger signal • Higher sensitivity • Real-time information • Simultaneous detection

for luciferase reporter assaysbrighter solutions

Thermo Scientific Luciferase Reporter Products

A bright new way for better reporter assays

Transcription Factors

Transcription

ReporterGene

mRNA

RNA Polymerase

AAAA(n)

Luciferase

Luciferase

Translation

+ Substrate

Luciferase Enzyme

Light

Light Signal = Luciferase Expression = Promoter Activity

Reporter Protein

Promoter

What is Bioluminescence?Bioluminescence is defined as the light- generating catalysis of a substrate by enzymes produced by living organisms. Application of this reaction in biological assays has become a powerful tool for mutational analysis of gene promoters, transcription factor dynamics, and protein-protein interactions.

Figure 1. Luciferase enzymes oxidize a luciferin substrate to produce light that can be measured with a luminometer. Luciferase-based reporter gene assays are widely used due to their ultrasensitive detection capacity and wide dynamic range. They involve placing a genetic regulatory element upstream of a luciferase gene and then transferring the resulting reporter construct into animal cells, plant cells or bacteria using a variety of methods. Expression of the luciferase reporter gene is then measured to quantify the activity of the regulatory element (cis-acting) or proteins (trans-acting) in the biological pathway affected by the regulatory element.

Luciferase Substrate + O2 ------------------------ Oxidized substrate + light

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Cypridina Gaussia Green Renilla Red Firefly

pMCS- Luc


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pUC Ori


Luc

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MCS

Lac01 TPS


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Select Luciferase

Vector

Add AssayReagents

Transfect Cells

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The new Thermo Scientific Pierce Luciferase Reporter Assays deliver greater assay sensitivity, signal stability and multiplexing capabilities. Some of these reporters also enable real-time kinetic studies without sacrificing valuable cell samples due to their secretory nature. The spectral resolution between our luciferase genes makes multiplexing easier.

We have developed five different luciferase reporter genes and a series of corresponding assay reagents to study gene regulation. These luciferase genes include Gaussia, Gaussia-Dura, Cypridina, Green Renilla and Red Firefly luciferases. These reporter genes can be used individually for single-reporter assays or in special combination for dual assays.

Gaussia, Gaussia-Dura and Cypridina luciferase contain a natural secretory signal peptide and are secreted into the cell culture media, allowing for live cell monitoring of reporter activity. The signal produced by these three secreted luciferases is considerably greater than signal from either Firefly or Renilla luciferases. Although more than 85% of the Gaussia, Gaussia-Dura and Cypridina luciferase is secreted, the remaining intracellular portion can be detected in lysates.

Green Renilla and Red Firefly luciferases lack a secretory peptide and are expressed and retained inside the cell. The Green Renilla luciferase is an intracellular protein with greater serum stability and higher light output compared

to native Firefly and Renilla luciferases. Consequently, Green Renilla reporter assays are more sensitive than assays using native Renilla luciferase. The Red Firefly luciferase is an intracellular protein with an emission peak shifted into the red spectrum with respect to the native Firefly luciferase. The spectral shift of Red Firefly allows multiplexing with Gaussia, Cypridina and Green Renilla (see Dual-Spectral Assays section).

Applications• Promoter studies for analyzing cis-acting 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

• Live-cell time-course studies

• Signal transduction pathway analysis

• Studying difficult-to-transfect cells, including stem cells and primary cells

• RNA splicing studies

Features Benefits

Higher sensitivity• 100-1,000X greater signal allows use of fewer cells

and decreases reagent consumption• Ideal for 384- and 1536-well plates

Better multiplexing capabilities

• Spectrally resolve multiple luciferases at the same time• One-step multiplex assays. No quenching step needed• Minimize assay variability

Real-time signal detection

• Non-destructive monitoring of luciferase activity in real time• Significantly cut down the cost and time of experimentation • Assay sensitivity at >3 orders of magnitude greater than

other luciferases

Table 1. Available plasmids and reporter gene characteristics.

Plasmid namePromoter used for expression Luciferase reporter

Is the reporter protein secreted in culture media? Which substrate is needed?

pMCS-Cypridina Luc NoneCypridina(62kDa)

Yes VargulinpCMV-Cypridina Luc CMV

pTK-Cypridina Luc TK

pMCS-Gaussia Luc NoneGaussia(22kDa)

Yes CoelenterazinepCMV-Gaussia Luc CMV

pTK-Gaussia Luc TK

pMCS-Gaussia-Dura Luc NoneGaussia-Dura

(22kDa)Yes CoelenterazinepCMV-Gaussia-Dura Luc CMV

pTK-Gaussia-Dura Luc TK

pMCS-Green Renilla Luc NoneGreen Renilla

(36kDa)No CoelenterazinepCMV-Green Renilla Luc CMV

pTK-Green Renilla Luc TK

pMCS-Red Firefly Luc NoneRed Firefly(62kDa)

No LuciferinpCMV-Red Firefly Luc CMV

pTK-Red Firefly Luc TK

Luciferase Reporter Assays

pMCS- Luc


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pUC Ori


Luc

SV

40 p

A

AmpR Ter

MCS

Lac01 TPS


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Select Luciferase

Vector

Add AssayReagents

Transfect Cells

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Figure 2. Thermo Scientific Pierce Flash Assay Reagents and Luciferases deliver the brightest light intensity. Panels A-C: Flash assay reagents were added to sample wells containing either media or lysates obtained from HEK293 cells (twenty-four hours post-transfection). Total light output (RLU) was captured by Thermo Scientific Varioskan Flash Luminometer (intergration time = 0.1 sec/reading). Panel D: Flash assay reagents were added to sample wells containing lysates obtained from HeLa cells (72 hours post-transfection). Total light output (RLU) was captured by Varioskan Flash Luminometer (intergration time = 1.0 sec/reading).

Figure 3. Panels A-D: Thermo Scientific Pierce Glow Assay Reagents result in higher light intensity and excellent signal stability. Glow assay reagents were added to sample wells containing either media or lysates obtained from HEK293 cells (twenty-four hours post-transfection). Total light output (RLU) was captured by Varioskan Luminometer (integration time = 1.0 sec/reading). Panel A: Pierce Cypridina Glow Assay. Panel B: Pierce Gaussia-Dura Glow Assay. Panel C: Pierce Green Renilla Glow Assay compared to Product R Renilla Luciferase Assay System. Panel D: Pierce Red Firefly Glow Assay compared to Product G Glow Luciferase Assay System.

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(media) (lysate) PromegaFirefly

Bright-Glo™

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Thermo Scientific Pierce Cypridina

Flash Assay

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Thermo Scientific Pierce Gaussia

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Thermo ScientificPierce FireflyFlash Assay

FireflyGene

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Thermo Scientific Pierce Green Renilla Glow Assay

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Thermo Scientific Pierce Gaussia

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FireflyGene

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Thermo Scientific Pierce Green Renilla Glow Assay

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Thermo Scientific Pierce Red Firefly Glow Assay

Luciferase assays can be read in either Flash or Glow format. The main difference between flash and glow luminescence lies in their different kinetic behavior. Flash reactions generate a transient luminescence signal after substrate is added, whereas glow reactions generate a stable luminescence signal for ~1 hour. Glow reactions are generally less sensitive than flash reactions.

Flash vs. Glow Assays

pMCS- Luc


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pUC Ori


Luc

SV

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AmpR Ter

MCS

Lac01 TPS

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Thermo Scientific Pierce Gaussia-Dura Glow Assay

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Select Luciferase

Vector

Add AssayReagents

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Thermo Scientific Luciferase Reporter Assays deliver high assay sensitivity. These reporters can be used to determine promoter activity, study signal transduction pathways in the cell or screen small molecules. Activity of our five different luciferase genes, Gaussia, Gaussia-Dura, Cypridina, Green Renilla and Red Firefly, can be determined individually using Pierce Flash or Glow Assay Reagent Kits. These single-reporter assays enable one to perform a nonradioactive assay for transcriptional activity of the regulatory elements. Three different luciferases with their specific substrates can be combined for multiplexed reporter assays.

Thermo Scientific Dual Luciferase Assays are highly sensitive flash-type assays for multiplexing. Typical dual assays include both an experimental luciferase activity and a second ‘control’ activity for normalization. The first luciferase activity reports specific experimental conditions and the second activity reflects transfection efficiency and cell viability.

Dual luciferase assays are ideal for analyzing: • Multiple regulatory elements simultaneously

• Multiple signaling pathways simultaneously

• More than one target per screen, including off-target effects

• Cross-talk between two or more pathways

We offer two unique types of dual assays for multiplexing:

A) Dual-spectral assay: This is a one-step procedure for spectrally resolving two different luciferases. By adding different substrates, luciferase activities can be measured simultaneously by filter-based detection in a single sample. This method eliminates the quenching step. We offer three spectral dual-assay systems: • Gaussia – Red Firefly Luciferases

• Cypridina – Red Firefly Luciferases

• Green Renilla – Red Firefly Luciferases

Figure 5. Dual-reporter assay procedure. BP = band pass. LP = long pass.

B) Dual-secreted assay: This is a two-step procedure for real-time monitoring of Gaussia- and Cypridina-secreted luciferases. It is a highly sensitive dual-assay system that enables kinetic measurement of luciferases from the same cell culture media, reducing sample handling time and decreasing assay variability. For more information about this assay, please contact us at [email protected].

Dual Assay

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Plate 104 cells/well in a96-well plate

Addtransfection

mixTransfect cells with

luciferase reporter plasmid

Treat cells as needed

Analyze results

Read luminescence signal

For analysis of intracellularluciferase, remove media and

lyse cells directly in wells

For analysis of secretedluciferase, transfer media

to a separate plate

Add luciferase assay reagent Add luciferase assay reagent

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options

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Add luciferaseassay reagent

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Plate 104 cellsper well in a 96-well plate

Transfect cells withluciferase reporter

plasmid

Remove mediaand lyse cells

directly in wells

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Addtransfection

mix

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Read luminescence signals

Light emitted at 480±20nm BP

Light emitted at 640nm LP

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Single Assay

Figure 4. Single-reporter assay procedure.

pMCS- Luc


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pUC Ori


Luc

SV

40 p

A

AmpR Ter

MCS

Lac01 TPS


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Select Luciferase

Vector

Add AssayReagents

Transfect Cells

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The pMCS-Luciferase vectors carry luciferase genes that are optimized for expression in mammalian cells, resulting in significantly improved light production. These vectors contain a multiple cloning site (MCS) for inserting a transcription regulatory element. A custom cloning service is available for inserting regulatory elements into the MCS of five different pMCS-luciferase vectors. Visit www.thermoscientific.com/pierce for more information.

The pCMV-Luciferase vectors have the luciferase gene under the CMV (Cytomegalovirus) promoter for constitutively high expression in mammalian cells. In pTK-Luciferase vectors, the luciferase gene is under the Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter for constitutively low-level expression in the mammalian cells. These pCMV and pTK plasmids can be used as normalization controls in transfection experiments in combination with other reporter vectors (Figure 6).

Highlights: • User-friendly MCS site for cloning regulatory element of choice

• CMV promoter for constitutively high luciferase expression

• TK promoter for constitutive low-level expression

• Transcription termination site (Ter), lac operator (Lac O1) and Transcriptional Pause Site (TPS) are used to minimize background by reducing transcriptional read-through

• Puromycin and ampicillin drug selection markers for mammalian cells and bacterial, respectively

• High-copy pVC bacterial DNA replication origin

Thermo Scientific TurboFect Transfection Reagent is ideal for transfection of a variety of cells, including primary and difficult-to-transfect cells. It is a sterile solution of a cationic polymer in water. Transfection can be performed in the presence or absence of serum. TurboFect™ Transfection Reagent demonstrates superior transfection efficiency and minimal toxicity when compared to lipid-based or other polymer-based transfection reagents. TurboFect Transfection Reagent has been validated with all our luciferase vectors.

Figure 7A. Successful transfection of many cell types with Thermo Scientific TurboFect Transfection Reagent. Comparison of transfection efficiency in various cell types transfected with a plasmid DNA encoding eGFP using TurboFect Transfection Reagent. Transfections (n = 2 or 3) were analyzed by flow cytometry.

Figure 7B. Better transfection with Thermo Scientific TurboFect Transfection Reagent. Comparison of transfection efficiency (left axis) and protein expression (right axis) in HepG2 cells transfected with a plasmid DNA encoding eGFP using TurboFect Transfection Reagent or products from other suppliers. Transfections were performed (n = 2 or 3) according to each manufacturer’s recommendations and analyzed by flow cytometry.

Select Luciferase Vector Transfect Cells

Figure 6. Each luciferase reporter gene is available in the pMCS, pCMV and pTK vectors.

pMCS- Luc


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pUC Ori


Luc

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MCS

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pCMV-LucpUC Ori

Amp R

SV40 pA Puromyc

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SV40

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Ter

PCMV Luc

BGH pA

Lac01 TPS

pTK-LucpUC Ori

Amp R

SV40 pA Puromyc

inR

SV40

Ori

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Ter

PTK Luc

BGH pA

Lac01 TPS

B50Calu1

CHOCOLO Cos7

HEK293

HeLa

HeLaS3

Hep2CHepG2

L929

MDCK

NIH3T

3

Raw264

Sp2/Ag14

WEHI

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pMCS- Luc


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Four luciferase reporter genes spectrally resolved at distinct wavelengths

Gaussia, Gaussia-Dura, and Cypridina luciferases are naturally secreted in the media. However, cell lysates still yield enough luciferase activity that can be used to measure with either flash or glow assay reagents. Gaussia luciferase performs best in flash assay mode, whereas Gaussia-Dura is optimized for glow assay mode. In dual-spectral assays, two luciferases are measured simultaneously in cell lysates adding a single reagent optimized for detection of two luciferases. Upon addition of this dual-assay reagent, the two luciferases emit spectrally resolvable luminescent light. These signals can be measured by filter-based detection in a single sample without any need for a quenching step.

Figure 8. Spectral emission profile for dual-spectral assays. Shaded ranges represent the filter sets used in product development experiments: 480±20nm band pass (BP) and 640nm long pass (LP).

Achieve quantitative results to accurately detect changes

The light output from a luciferase reaction is measured using a luminometer. Results are provided as a Relative Light Unit (RLU) value, so variations can be easily identified and quantified.

No filter is needed for single assays because all generated photons are measured. For dual assays, filters are needed to selectively read light emission at wavelengths specified for the luciferase gene used (Table 2). Our spectral dual assays involve measuring two luciferase genes with emission peaks that are easily resolvable with conventional filters (Figure 8).

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Cypridina Red Firefly

Add Assay Reagents Read Signal

Table 2. Luciferase/substrate combinations for flash and glow assays.

Single assay Isolate luc from Substrate Assay type Recommended filters

Cypridina Media or lysate Vargulin Flash, Glow

No filter needed.Collect total light output.

Gaussia Media or lysate Colenterazine Flash

Gaussia-Dura Media or lysate Colenterazine Glow

Green Renilla Lysate Colenterazine Flash, Glow

Red Firefly Lysate D-Luciferin Flash, Glow

Spectral dual assay Substrate Assay type Recommended filters

Cypridina/Red Firefly Lysate Vargulin & D-Luciferin Dual Assay 480±20nm & 640nm long pass

Gaussia/Red Firefly Lysate Coelenterazine & D-Luciferin Dual Assay 480±20nm & 640nm long pass

Green Renilla/Red Firefly Lysate Coelenterazine & D-Luciferin Dual Assay 525±20nm & 640nm long pass

Dual-secreted assay Substrate Assay type Recommended filters

Cypridina/Gaussia Media Vargulin/Colenterazine Dual Assay Not required. Collect total light output.

pMCS- Luc


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pUC Ori


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MCS

Lac01 TPS


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Product B Bright Glow Assay System

Gaussia (lysate)Cypridina (lysate)

Green Renilla

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Figure 12 Panel A. Thermo Scientific Pierce Gaussia Luciferase and optimized assay reagents allow the luciferase signal to be detected in difficult-to-transfect cells. Thermo Scientific pCMV-Gaussia Plasmid (100ng) was transfected in Jurkat suspension cells using TurboFect Transfection Reagent. Transfection efficiency of <1% was observed

by FACS analysis using a CMV-GFP control plasmid. Twenty-four hours after the transfection, 20μL of cell culture media was used to measure luciferase activity in the flash assay reagent.

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Thermo Scientific PierceGaussia Flash Assay

Product F FireflyGlow Assay

Figure 12 Panel B. Thermo Scientific Luciferase Assay Systems show higher signal outputs than Product F Firefly Glow Assay in difficult-to-transfect Jurkat cells. Four different Thermo Scientific pCMV-Luciferase plasmids were individually transfected in Jurkat

suspension cells by using TurboFect Transfection Reagent. Twenty-four hours after the transfection, Jurkat suspension cells were spun down by centrifugation with 1000xg for 5 min. The media was transferred to a fresh tube. Cell culture media (20μL) was used to measure the luciferase activity in the Thermo Scientific Luciferase Flash Assay Reagents. The cells were lysed with 100μL 1X Pierce Luciferase Cell Lysis Buffer.

Thermo ScientificLuciferase Assay System

Product F FireflyGlow Assay

Cyp (media)Cyp (lysate)

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Thermo Scientific PierceDual Assay Systems

Product D DualAssay System

Figure 14. Thermo Scientific Dual Assay delivers superior results.Thermo Scientific Spectral Dual Assay was compared with the Product D Dual Assay System. Fifty nanograms each of pCRE-Luc reporter plasmids and pCMV-Luc control plasmids were transfected HEK293 cells by using TurboFect Transfection Reagent. Twenty-four hours after the transfection, the culture media was removed and replenished with fresh media containing 30μM forskolin to activate CRE promoter. Three hours after activation, cells were lysed with 100μL 1X Pierce Luciferase Cell Lysis Buffer. Cell lysate (20μL) was used to measure luciferase activity in either Thermo Scientific Gaussia/Firefly Luciferase Assay Kit or Product D Dual Assay System.

MultiplexingHigher Sensitivity

Figure 11. Thermo Scientific Pierce Luciferases can be detected in as few as 1000 cells. Thermo Scientific pCMV-Luc plasmids (100ng) were transfected in HEK293 cells by using TurboFect Transfection Reagent. Twenty-four hours after the transfection, the cells were lysed with 100μL 1X Pierce Luciferase Cell Lysis Buffer. Cell lysate (20μL) was used to measure luciferase activity in the flash assay reagents.

Figure 13. Real-time monitoring of CREB and NFκB signal transduction pathways using secreted luciferase reporters. Three plasmids, pCRE-Gaussia, NFκB-Cypridina, and CMV-Red Firefly, were co-transfected into HEK293 cells using TurboFect Transfection Reagent. Twenty-four hours after the transfection, the cells were treated with forskolin for 3 hours to increase intracellular cAMP level and activate the Gaussia luciferase reporter fused to CRE promoter. The secreted Gaussia in media was measured with Pierce Gaussia Flash Assay Kit. Simultaneously, the effect of cAMP accumulation on NFκB survival pathway was monitored by using NFκB-Cypridina as second secreted reporter in real time without sacrificing the cells. After 7 hours of forskolin stimulation, the cells were lysed and CMV-Red Firefly activity was measured using the Pierce Firefly Flash Assay Kit. Signals obtained were used to normalize the Cypridina and Gaussia luciferase signals.

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pMCS- Luc


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pUC Ori


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Need a larger quantity?Call our Bulk and Custom department at 1-800-874-3723

or +1 815-968-0747 ext. 300 for pricing and information. Or, visit www.thermoscientific.com/protein-custom

Bulk-sizePackagesAvailable

Product # Description Pkg. Size

PI-16146 pMCS-Gaussia Luc, 0.5μg/μL 10µg

PI-16147 pCMV-Gaussia Luc, 0.5μg/μL 10µg

PI-16148 pTK-Gaussia Luc, 0.5μg/μL 10µg

PI-16149 pMCS-Cypridina Luc, 0.5μg/μL 10µg

PI-16150 pCMV-Cypridina Luc, 0.5μg/μL 10µg

PI-16151 pTK-Cypridina Luc, 0.5μg/μL 10µg

PI-16152 pMCS-Green Renilla Luc, 0.5μg/μL 10µg

PI-16153 pCMV-Green Renilla Luc, 0.5μg/μL 10µg

PI-16154 pTK-Green Renilla Luc, 0.5μg/μL 10µg

PI-16155 pMCS-Red Firefly Luc, 0.5μg/μL 10µg

PI-16156 pCMV-Red Firefly Luc, 0.5μg/μL 10µg

PI-16157 pTK-Red Firefly Luc, 0.5μg/μL 10µg

PI-16190 pMCS-Gaussia-Dura Luc, 0.5μg/μL 10µg

PI-16191 pCMV-Gaussia-Dura Luc, 0.5μg/μL 10µg

PI-16192 pTK-Gaussia-Dura Luc, 0.5μg/μL 10µg

PI-16189 Pierce Luciferase Cell Lysis Buffer (2X) 250mL

PI-R0533 TurboFect Transfection Reagent 200µL

PI-R0534 TurboFect Transfection Reagent 5 x 200µL

PI-R0532 TurboFect Transfection Reagent 5 x 1mL

Single Assay Kits Pierce Gaussia Luciferase Flash Assay Kits Gaussia Flash Assay Buffer 100X Coelenterazine Luciferase Cell Lysis Buffer (2X)

PI-16158 Gaussia Luciferase Flash Assay Kit 100-rxn kit

PI-16159 Gaussia Luciferase Flash Assay Kit 1000-rxn kit

Pierce Gaussia-Dura Luciferase Glow Assay Kits Gaussia Glow Assay Buffer 100X Coelenterazine Luciferase Cell Lysis Buffer (2X)

PI-16160 Gaussia Luciferase Glow Assay Kit 100-rxn kit

PI-16161 Gaussia Luciferase Glow Assay Kit 1000-rxn kit

Pierce Renilla Luciferase Flash Assay Kits Renilla Flash Assay Buffer 100X Coelenterazine Luciferase Cell Lysis Buffer (2X)

PI-16164 Renilla Luciferase Flash Assay Kit 100-rxn kit

PI-16165 Renilla Luciferase Flash Assay Kit 1000-rxn kit

Pierce Renilla Luciferase Glow Assay Kits Renilla Glow Assay Buffer 100X Coelenterazine Luciferase Cell Lysis Buffer (2X)

PI-16166 Renilla Luciferase Glow Assay Kit 100-rxn kit

PI-16167 Renilla Luciferase Glow Assay Kit 1000-rxn kit

Product # Description Pkg. Size

Pierce Cypridina Luciferase Flash Assay Kits Cypridina Flash Assay Buffer 100X Vargulin Luciferase Cell Lysis Buffer (2X)

PI-16168 Cypridina Luciferase Flash Assay Kit 100-rxn kit

PI-16169 Cypridina Luciferase Flash Assay Kit 1000-rxn kit

Pierce Cypridina Luciferase Glow Assay Kits Cypridina Glow Assay Buffer 100X Vargulin Luciferase Cell Lysis Buffer (2X)

PI-16170 Cypridina Luciferase Glow Assay Kit 100-rxn kit

PI-16171 Cypridina Luciferase Glow Assay Kit 1000-rxn kit

Pierce Firefly Luciferase Flash Assay Kits Firefly Flash Assay Buffer D-Luciferin Luciferase Cell Lysis Buffer (2X)

PI-16174 Firefly Luciferase Flash Assay Kit 100-rxn kit

PI-16175 Firefly Luciferase Flash Assay Kit 1000-rxn kit

Pierce Firefly Luciferase Glow Assay Kits Firefly Glow Assay Buffer D-Luciferin Luciferase Cell Lysis Buffer (2X)

PI-16176 Firefly Luciferase Glow Assay Kit 100-rxn kit

PI-16177 Firefly Luciferase Glow Assay Kit 1000-rxn kit

PI-16180 Firefly Signal Enhancer, (100X), 0.5μL 0.5mL

Dual Assay KitsPierce Gaussia-Firefly Luciferase Dual Assay Kits Gaussia-Firefly Dual Assay Buffer 100X Coelenterazine D-Luciferin Luciferase Cell Lysis Buffer (2X)

PI-16181 Gaussia-Firefly Luciferase Dual Assay Kit 100-rxn kit

PI-16182 Gaussia-Firefly Luciferase Dual Assay Kit 1000-rxn kit

Pierce Cypridina-Firefly Luciferase Dual Assay Kits Cypridina-Firefly Dual Assay Buffer 100X Vargulin D-Luciferin Luciferase Cell Lysis Buffer (2X)

PI-16183 Cypridina-Firefly Luciferase Dual Assay Kit 100-rxn kit

PI-16184 Cypridina-Firefly Luciferase Dual Assay Kit 1000-rxn kit

Pierce Renilla-Firefly Luciferase Dual Assay Kits Renilla-Firefly Dual Assay Buffer 100X Coelenterazine D-Luciferin Luciferase Cell Lysis Buffer (2X)

PI-16185 Renilla-Firefly Luciferase Dual Assay Kit 100-rxn kit

PI-16186 Renilla-Firefly Luciferase Dual Assay Kit 1000-rxn kit

Ordering Information

pMCS- Luc


17131319253137

pUC Ori


Luc

SV

40 p

A

AmpR Ter

MCS

Lac01 TPS


17131319253137

Select Luciferase

Vector

Add AssayReagents

Transfect Cells

ABCDEFGH

1 2 3 4 5 6 7 8 9 10 11 12


1 2 3 4 5 6 7 8 9 10 11 12

400 450 500 550 600 650 700

Wavelength (nm)

Rela

tive

Ligh

t Uni

ts (R

LU),

Nor

mal

ized


www.thermoscientific.com/pierce

In the United States:For customer service, call 1-800-766-7000.To fax an order, use 1-800-926-1166.To order online: www.fishersci.com

In Canada:For customer service, call 1-800-234-7437.To fax an order, use 1-800-463-2996.To order online: www.fishersci.ca

BN01121213 01/12 Printed in the U.S.

www.thermoscientific.com/pierce© 2012 Thermo Fisher Scientific Inc. All rights reserved. These products are supplied for laboratory or manufacturing applications only. All trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries.

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