Secrete-Pair Dual Luminescence Assay Kit

Introduction

Secrete-Pair™ Gaussia Luciferase Dual and Single Luminescence Assay Kits

GeneCopoeia's Secrete-Pair™ Dual Luminescence Assay Kit is designed to analyze the activities of Gaussia Luciferase (GLuc) and Secreted Alkaline Phosphatase (SEAP) in a dual-reporter system. Both GLuc and SEAP are secreted reporter proteins, permitting detection without cell lysis. Secrete-Pair measures dual reporter signals and allows transfection normalization. A single GLuc reporter system is also available.

 

Advantages

• Live cell assays. Secreted GLuc and SEAP make cell lysis unnecessary. The same sample can be assayed multiple times for different time points, environmental conditions, etc., reducing sample-to-sample variation. Ideal for high-throughput applications.

• Dual-reporter detection. SEAP allows normalization of GLuc signal for greater accuracy.

• Sensitive and robust system. GLuc is 1,000 times more sensitive than firefly and Renilla luciferases.

• Flexible assay conditions. Two robust buffer conditions are provided for GLuc assays: 1) Buffer for greater stability retains more than 90% of signal within the first 10 minutes, extending the half-life of luminescence to 30 minutes; 2) Buffer for higher sensitivity for detecting low GLuc expression.

• Clone and vector compatibility. Compatible with GeneCopoeia's GLuc-ON™ promoter reporter clones, miTarget™ miRNA target clones, GLuc-ON™ Transcriptional Response Element (TRE) clones, and cloning vectors.

 

To Order

Secrete-Pair™ kits

 

Buy Catalog# Product Description Price
LF031 Secrete-Pair Dual Luminescence Assay Kit (100 rxns) Detects Gaussia luciferase (GLuc) and secreted alkaline phosphatase (SEAP) $139

LF032

30% off

Secrete-Pair Dual Luminescence Assay Kit (300 rxns) $379

LF033

30% off

Secrete-Pair Dual Luminescence Assay Kit (1000 rxns) $959
LF061 Secrete-Pair Gaussia Luciferase Assay Kit (100 rxns) Detects Gaussia luciferase (GLuc) $77

LF062

30% off

Secrete-Pair Gaussia Luciferase Assay Kit (1000 rxns) $399

 

  User manual MSDS

 

Cloning vectors 

GeneCopoeia also offers Gaussia luciferase (Gluc) reporter vectors shown below. Click on the name to view the vector map. (Discounted price – $199)

Buy Catalog# Product Description Promoter Reporter gene Tracking gene Selection marker Size Price
ZX101 pEZX-GN01 Gaussia luciferase (Gluc) reporter cloning vector N/A* Gluc N/A* Puromycin (10µg) $375
ZX102 pEZX-GN03 Gaussia luciferase (Gluc) reporter cloning vector miniCMV Gluc N/A* Puromycin (10µg) $375
ZX103 pEZX-GA01 Gaussia luciferase (Gluc) and secreted alkaline phosphatase (SEAP) reporter cloning vector N/A* Gluc SEAP Puromycin (10µg) $495
ZX104 pEZX-GA02 Gaussia luciferase (Gluc) and secreted alkaline phosphatase (SEAP) reporter cloning vector SV40 Gluc SEAP Neomycin (10µg) $495
ZX105 pEZX-GA03 Gaussia luciferase (Gluc) and secreted alkaline phosphatase (SEAP) reporter cloning vector miniCMV Gluc SEAP Puromycin (10µg) $495
ZX106 pEZX-LvGN01 Gaussia luciferase (Gluc) reporter cloning vector (lentiviral) N/A* Gluc N/A* Puromycin (10µg) $375
ZX107 pEZX-LvGA01 Gaussia luciferase (Gluc) and secreted alkaline phosphatase (SEAP) reporter cloning vector (lentiviral) N/A* Gluc SEAP Puromycin (10µg) $495

* A separate vector is available for SEAP expression

 

 

Related products

SEAP normalization control clone

Buy Cat.No. Product name Price
SEAP-PA01 Secreted alkaline phosphatase (SEAP) Expression clone $395


 
SEAP-LvPA02 Secreted alkaline phosphatase (SEAP) Expression clone(lentiviral vector) $595

 

GLuc positive control clone

Buy Cat.No. Product name Price
GAPDH-PG02 GAPDH positive control clone $375

 

EndoFectin™ transfection reagents

Buy Cat.No. Product name Price Data sheets
EFM1004-01 EndoFectin™ Max transfection reagent (1 mL) $295 Download
EFM1004-02 EndoFectin™ Max transfection reagent (3 mL) $589
EFL1001-01 EndoFectin™ Lenti transfection reagent (1 mL) $245 Download
EFL1001-02 EndoFectin™ Lenti transfection reagent (3 mL) $489
EF005 EndoFectin™ HepG2 transfection reagent (1 mL) $355 Download
EF006 EndoFectin™ HepG2 transfection reagent (3 mL) $705
EF007 EndoFectin™ Expi293 transfection reagent (1 mL) $320  
EF008 EndoFectin™ Expi293 transfection reagent (3 mL) $640  

 

Additional related clones

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Additional related reagents

 

Protocol and Data

Protocol Overview

Figure 1. General workflow for the Secrete-Pair™ Dual Luminesence Assay System.

 

Dual-reporter system

The Secrete-Pair™ Dual Luminescence Assay System is ideally suited for dual-reporter vectors and clones, such as those available for the GLuc-ON™ promoter clones and miTarget™ microRNA target clones. The secondary reporter, secreted Alkaline Phosphatase (SEAP), serves as an internal control. The dual-reporter system enables transfection normalization for accurate cross-sample comparison (Figure 2).

 

Figure 2. Normalized promoter activities in H1B1B and HEK293T cells. Dual-reporter promoter clones or controls were transfected into two cell lines in duplicates. Samples were analyzed 24 hrs (HEK293T) and 48 hrs (H1B1B) after transfection. NEG (containing non-promoter sequence) and EMPTY (no promoter in the vector) are negative controls.

 

Two buffer conditions are provided in the kit for GLuc assays depending on the application. Buffer GL-S contains a stabilizer and can be used for stabilized activity by overcoming the quick decay of the GLuc signal. When higher signal intensities are needed, Buffer GL-H is used instead (Figure 3).

 

Figure 3. GLuc and SEAP assays. Cell culture medium was collected from cells transfected with wild-type (wt) GLuc- SEAP dual-reporter clone. 10 μl of the medium was used in each assay. At the beginning, the GLuc activity in Buffer GL- H is about 4-6 times higher than that in Buffer GL-S, then quickly decays. The GLuc activity in Buffer GL-S, however, is much more stable.

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2017

2016