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Cell biology assays

Luciferase reporters—Ready-To-Glow secreted luciferase assay

The Ready-To-Glow Secreted Luciferase Reporter Assay is based on the secreted Metridia luciferase reporter. It combines the advantages of a live-cell assay with the sensitivity of an enzyme-based assay.

Our Ready-To-Glow MetLuc Monoclonal Antibody makes it easy to detect Metridia luciferase expressed by mammalian cells, as well as recombinant Metridia luciferase, in amounts down to 20 ng using western blot analysis.

Benefits of using a live-cell assay

	Ready-To-Glow Secreted Luciferase Reporter Assay	Firefly and Renilla-Based Reporter Assays
One-step protocol, without cell lysis	Yes. Reporter molecule is naturally secreted into the media and can be assayed without lysing cells.	No.
2–4 fold higher signal than Renilla or firefly assays	Yes.	No.
Multiple data points per well	Yes. Single well can be used for multiple time points, as there is no cell lysis.	No.
Multiple compounds/trigger testing on same cells (single transfection)	Yes.	No.
Removal of background signal	Yes. Background signal can be reset to zero by removing and replacing the media	No.

Dual secreted reporter assay—luciferase & SEAP

The Ready-To-Glow Dual Secreted Reporter Assay has two live-cell reporters that do not require cell lysis. In addition to the secreted luciferase assay, it includes a secreted version of alkaline phosphatase.

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Cat. #	Product	Size	Price
631781	Ready-To-Glow™ MetLuc Monoclonal Antibody	50 ug	USD $618.00
The Ready-To-Glow MetLuc Monoclonal Antibody recognizes Metridia luciferase expressed by mammalian cells, as well as recombinant Metridia luciferase, in amounts down to 20 ng using Western blot analysis. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components You May Also Like Image Data Back 631781: Ready-To-Glow MetLuc Monoclonal Antibody
631740	Ready-To-Glow™ Automation Kit	5,000 Rxns	USD $2076.00
Our Ready-To-Glow Automation Kit is a complete detection system for analysis of gene activity via plate readers with automated injection systems. It is designed for use with a transcription reporter system based on Metridia luciferase activity in the supernatant of transfected cells. In the presence of secreted luciferase in the sample, a nonluminescent substrate is converted to a luminescent compound. The intensity of bioluminescence is directly proportional to the amount of luciferase in the sample. The kit includes substrate buffer, reaction buffer, and a lyophilized secreted luciferase substrate. It has been optimized to work with our Ready-To-Glow Secreted Luciferase Vector Kit (Cat. No. 631729). Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components You May Also Like Image Data Back Limit of detection for recombinant Metridia luciferase protein *Limit of detection for recombinant Metridia* luciferase protein.** The indicated amounts of recombinant Metridia luciferase (fg/well) were spiked into DMEM containing 10% FBS. Metridia luciferase activity was measured in a 96-well format on the SpectraMax L. The limit of detection was determined to be 2 fg/well (40 fg/ml). Back High signal intensity and stability using secreted Metridia luciferase *High signal intensity and stability using secreted Metridia* luciferase.** CHO cells were plated into 96-well plates and transiently transfected with CMV-driven constructs encoding non-secreted firefly luciferase, non-secreted Renilla luciferase, and secreted Metridia luciferase. 24 hr after transfection, luciferase activity in equivalent samples was analyzed by addition of the recommended substrate. The signal was measured at different timepoints over a period of 45 min. neg = negative control. Back Monitoring promoter activation using the sequence-optimized secreted Metridia luciferase reporter *Monitoring promoter activation using the sequence-optimized secreted Metridia* luciferase reporter.** HeLa cells were transiently transfected with a vector construct containing the NFkB response element driving the expression of sequence-optimized secreted Metridia luciferase. 24 hr after transfection, the media was removed and replaced by media with or without TNF-α (100 ng/ml) to activate the NFkB signal transduction pathway. Six hr after addition of TNF-α, samples of the media were removed and analyzed for Metridia luciferase activity. The fold induction was calculated for different time points following the addition of substrate. Back Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System. Back Linear range of detection of Ready-To-Glow secreted Metridia luciferase from media supernatants *Linear range of detection of Ready-To-Glow secreted Metridia* luciferase from media supernatants.** HEK 293 cells and CHO cells were transiently transfected with the pMetLuc-Control vector. Media samples were collected from the transfected cells and 1:10 serial dilutions were performed in DMEM containing 10% FBS, to the concentrations noted. Ready-To-Glow Metridia luciferase activity was measured according to the luciferase assay protocol for either 96-well (Panel A) or 384-well (Panel B) plates. Back Use of secreted Metridia luciferase in a high-throughput screening (HTS) application *Use of secreted Metridia* luciferase in a high-throughput screening (HTS) application.** The figure shows a screen shot from a CCD camera of a 1,536-well plate containing stable CHO cells transfected with a forskolin-responsive Metridia luciferase gene (pASM-Lu164). 300 cells/well were plated in a 1,536-well microtiter plate. Cells were incubated with increasing concentrations of forskolin and the plate was incubated for 4 hr at 37° C. Metridia substrate was added in increasing concentrations and the plate was visualized using a CCD camera system (integration time: 60 sec). (Courtesy of Bayer Health Care, Germany). This pseudocolor image reflects luminescence intensity. Yellow regions are the brightest, while black regions are the least bright. Back 631740: Ready-To-Glow Automation Kit
631734	Ready-To-Glow™ Dual Secreted Reporter Assay	500 Rxns	USD $970.00
The Ready-To-Glow Dual Secreted Reporter Assay contains the buffers and substrates for use with the vectors contained in the Ready-To-Glow Dual Secreted Reporter Vector Kit (Cat. No. 631735). This assay allows you to monitor the activity of two promoters simultaneously. The promoters are monitored by testing the media from cells that have been transfected with the two reporter vectors, which express luciferase and SEAP respectively. The assay allows you to monitor promoter activity over time, because samples of the cell media can be taken repeatedly and tested without lysing cells. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components You May Also Like Image Data Resources Back Similar secretion kinetics of Metridia secreted luciferase and SEAP enable accurate comparisons of the relative timing of promoter activity *Similar secretion kinetics of Metridia* secreted luciferase and SEAP enable accurate comparisons of the relative timing of promoter activity. HeLa cells were plated into 6-well plates and transiently transfected with either the pMetLuc-Control vector or the pSEAP-Control vector. Media samples from the transfected cells were collected from 3 wells at each time point by removing enough media to run either the luciferase or the SEAP assay. Each sample was tested in triplicate, using a white-bottom 96-well microtiter plate on a Turner BioSystems Veritas Luminometer. Back Monitoring activation of two promoters simultaneously Monitoring activation of two promoters simultaneously.** HEK 293 cells cotransfected with pNF kappa B-TA-MetLuc and pCRE-SEAP constructs were treated with fresh media alone or with media containing either 1,000 ng/ml TNF-alpha or 10 µM forskolin. Samples of culture supernatant were collected 7 hr later and assayed using a BD Monolight 3096 Luminometer. Back 631734: Ready-To-Glow Dual Secreted Reporter Assay
631728	Ready-To-Glow™ Secreted Luciferase Reporter Assay	1,000 Rxns	USD $642.00
Our Ready-To-Glow Secreted Luciferase Reporter Assay is a complete system designed for use in a transcription reporter system based on luciferase activity in the supernatant of transfected cells. In the presence of secreted luciferase in the sample, a nonluminescing substrate is converted to a luminescent compound. The intensity of bioluminescence is directly proportional to the amount of luciferase activity. The kit, which includes a substrate buffer, reaction buffer, and a lyophilized secreted luciferase substrate, is available in three reaction sizes. Our Ready-To-Glow Secreted Luciferase Reporter Assay has been customized to work optimally with our Secreted Luciferase Vector Kits. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components You May Also Like Image Data Resources Back Limit of detection for recombinant Metridia luciferase protein *Limit of detection for recombinant Metridia* luciferase protein.** The indicated amounts of recombinant Metridia luciferase (fg/well) were spiked into DMEM containing 10% FBS. Metridia luciferase activity was measured in a 96-well format on the SpectraMax L. The limit of detection was determined to be 2 fg/well (40 fg/ml). Back High signal intensity and stability using secreted Metridia luciferase *High signal intensity and stability using secreted Metridia* luciferase.** CHO cells were plated into 96-well plates and transiently transfected with CMV-driven constructs encoding non-secreted firefly luciferase, non-secreted Renilla luciferase, and secreted Metridia luciferase. 24 hr after transfection, luciferase activity in equivalent samples was analyzed by addition of the recommended substrate. The signal was measured at different timepoints over a period of 45 min. neg = negative control. Back Monitoring promoter activation using the sequence-optimized secreted Metridia luciferase reporter *Monitoring promoter activation using the sequence-optimized secreted Metridia* luciferase reporter.** HeLa cells were transiently transfected with a vector construct containing the NFkB response element driving the expression of sequence-optimized secreted Metridia luciferase. 24 hr after transfection, the media was removed and replaced by media with or without TNF-α (100 ng/ml) to activate the NFkB signal transduction pathway. Six hr after addition of TNF-α, samples of the media were removed and analyzed for Metridia luciferase activity. The fold induction was calculated for different time points following the addition of substrate. Back Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System. Back Linear range of detection of Ready-To-Glow secreted Metridia luciferase from media supernatants *Linear range of detection of Ready-To-Glow secreted Metridia* luciferase from media supernatants.** HEK 293 cells and CHO cells were transiently transfected with the pMetLuc-Control vector. Media samples were collected from the transfected cells and 1:10 serial dilutions were performed in DMEM containing 10% FBS, to the concentrations noted. Ready-To-Glow Metridia luciferase activity was measured according to the luciferase assay protocol for either 96-well (Panel A) or 384-well (Panel B) plates. Back Use of secreted Metridia luciferase in a high-throughput screening (HTS) application *Use of secreted Metridia* luciferase in a high-throughput screening (HTS) application.** The figure shows a screen shot from a CCD camera of a 1,536-well plate containing stable CHO cells transfected with a forskolin-responsive Metridia luciferase gene (pASM-Lu164). 300 cells/well were plated in a 1,536-well microtiter plate. Cells were incubated with increasing concentrations of forskolin and the plate was incubated for 4 hr at 37° C. Metridia substrate was added in increasing concentrations and the plate was visualized using a CCD camera system (integration time: 60 sec). (Courtesy of Bayer Health Care, Germany). This pseudocolor image reflects luminescence intensity. Yellow regions are the brightest, while black regions are the least bright. Back 631728: Ready-To-Glow Secreted Luciferase Reporter Assay
631727	Ready-To-Glow™ Secreted Luciferase Reporter Assay	500 Rxns	USD $459.00
Our Ready-To-Glow Secreted Luciferase Reporter Assay is a complete system designed for use in a transcription reporter system based on luciferase activity in the supernatant of transfected cells. In the presence of secreted luciferase in the sample, a nonluminescing substrate is converted to a luminescent compound. The intensity of bioluminescence is directly proportional to the amount of luciferase activity. The kit, which includes a substrate buffer, reaction buffer, and a lyophilized secreted luciferase substrate, is available in three reaction sizes. Our Ready-To-Glow Secreted Luciferase Reporter Assay has been customized to work optimally with our Secreted Luciferase Vector Kits. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components You May Also Like Image Data Resources Back 631727: Ready-To-Glow Secreted Luciferase Reporter Assay Back Limit of detection for recombinant Metridia luciferase protein *Limit of detection for recombinant Metridia* luciferase protein.** The indicated amounts of recombinant Metridia luciferase (fg/well) were spiked into DMEM containing 10% FBS. Metridia luciferase activity was measured in a 96-well format on the SpectraMax L. The limit of detection was determined to be 2 fg/well (40 fg/ml). Back High signal intensity and stability using secreted Metridia luciferase *High signal intensity and stability using secreted Metridia* luciferase.** CHO cells were plated into 96-well plates and transiently transfected with CMV-driven constructs encoding non-secreted firefly luciferase, non-secreted Renilla luciferase, and secreted Metridia luciferase. 24 hr after transfection, luciferase activity in equivalent samples was analyzed by addition of the recommended substrate. The signal was measured at different timepoints over a period of 45 min. neg = negative control. Back Monitoring promoter activation using the sequence-optimized secreted Metridia luciferase reporter *Monitoring promoter activation using the sequence-optimized secreted Metridia* luciferase reporter.** HeLa cells were transiently transfected with a vector construct containing the NFkB response element driving the expression of sequence-optimized secreted Metridia luciferase. 24 hr after transfection, the media was removed and replaced by media with or without TNF-α (100 ng/ml) to activate the NFkB signal transduction pathway. Six hr after addition of TNF-α, samples of the media were removed and analyzed for Metridia luciferase activity. The fold induction was calculated for different time points following the addition of substrate. Back Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System. Back Linear range of detection of Ready-To-Glow secreted Metridia luciferase from media supernatants *Linear range of detection of Ready-To-Glow secreted Metridia* luciferase from media supernatants.** HEK 293 cells and CHO cells were transiently transfected with the pMetLuc-Control vector. Media samples were collected from the transfected cells and 1:10 serial dilutions were performed in DMEM containing 10% FBS, to the concentrations noted. Ready-To-Glow Metridia luciferase activity was measured according to the luciferase assay protocol for either 96-well (Panel A) or 384-well (Panel B) plates. Back Use of secreted Metridia luciferase in a high-throughput screening (HTS) application *Use of secreted Metridia* luciferase in a high-throughput screening (HTS) application.** The figure shows a screen shot from a CCD camera of a 1,536-well plate containing stable CHO cells transfected with a forskolin-responsive Metridia luciferase gene (pASM-Lu164). 300 cells/well were plated in a 1,536-well microtiter plate. Cells were incubated with increasing concentrations of forskolin and the plate was incubated for 4 hr at 37° C. Metridia substrate was added in increasing concentrations and the plate was visualized using a CCD camera system (integration time: 60 sec). (Courtesy of Bayer Health Care, Germany). This pseudocolor image reflects luminescence intensity. Yellow regions are the brightest, while black regions are the least bright.
631726	Ready-To-Glow™ Secreted Luciferase Reporter Assay	100 Rxns	USD $133.00
Our Ready-To-Glow Secreted Luciferase Reporter Assay is a complete system designed for use in a transcription reporter system based on luciferase activity in the supernatant of transfected cells. In the presence of secreted luciferase in the sample, a nonluminescing substrate is converted to a luminescent compound. The intensity of bioluminescence is directly proportional to the amount of luciferase activity. The kit, which includes a substrate buffer, reaction buffer, and a lyophilized secreted luciferase substrate, is available in three reaction sizes. Our Ready-To-Glow Secreted Luciferase Reporter Assay has been customized to work optimally with our Secreted Luciferase Vector Kits. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components You May Also Like Image Data Resources Back Limit of detection for recombinant Metridia luciferase protein *Limit of detection for recombinant Metridia* luciferase protein.** The indicated amounts of recombinant Metridia luciferase (fg/well) were spiked into DMEM containing 10% FBS. Metridia luciferase activity was measured in a 96-well format on the SpectraMax L. The limit of detection was determined to be 2 fg/well (40 fg/ml). Back High signal intensity and stability using secreted Metridia luciferase *High signal intensity and stability using secreted Metridia* luciferase.** CHO cells were plated into 96-well plates and transiently transfected with CMV-driven constructs encoding non-secreted firefly luciferase, non-secreted Renilla luciferase, and secreted Metridia luciferase. 24 hr after transfection, luciferase activity in equivalent samples was analyzed by addition of the recommended substrate. The signal was measured at different timepoints over a period of 45 min. neg = negative control. Back Monitoring promoter activation using the sequence-optimized secreted Metridia luciferase reporter *Monitoring promoter activation using the sequence-optimized secreted Metridia* luciferase reporter.** HeLa cells were transiently transfected with a vector construct containing the NFkB response element driving the expression of sequence-optimized secreted Metridia luciferase. 24 hr after transfection, the media was removed and replaced by media with or without TNF-α (100 ng/ml) to activate the NFkB signal transduction pathway. Six hr after addition of TNF-α, samples of the media were removed and analyzed for Metridia luciferase activity. The fold induction was calculated for different time points following the addition of substrate. Back Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System. Back Linear range of detection of Ready-To-Glow secreted Metridia luciferase from media supernatants *Linear range of detection of Ready-To-Glow secreted Metridia* luciferase from media supernatants.** HEK 293 cells and CHO cells were transiently transfected with the pMetLuc-Control vector. Media samples were collected from the transfected cells and 1:10 serial dilutions were performed in DMEM containing 10% FBS, to the concentrations noted. Ready-To-Glow Metridia luciferase activity was measured according to the luciferase assay protocol for either 96-well (Panel A) or 384-well (Panel B) plates. Back Use of secreted Metridia luciferase in a high-throughput screening (HTS) application *Use of secreted Metridia* luciferase in a high-throughput screening (HTS) application.** The figure shows a screen shot from a CCD camera of a 1,536-well plate containing stable CHO cells transfected with a forskolin-responsive Metridia luciferase gene (pASM-Lu164). 300 cells/well were plated in a 1,536-well microtiter plate. Cells were incubated with increasing concentrations of forskolin and the plate was incubated for 4 hr at 37° C. Metridia substrate was added in increasing concentrations and the plate was visualized using a CCD camera system (integration time: 60 sec). (Courtesy of Bayer Health Care, Germany). This pseudocolor image reflects luminescence intensity. Yellow regions are the brightest, while black regions are the least bright. Back 631726: Ready-To-Glow Secreted Luciferase Reporter Assay

Overview

Secreted luciferase assay protocol eliminates cell lysis
2–4 fold higher signal than firefly or Renilla luciferase
Single transfection allows multiple time points from same cell
Dual, live-cell assay includes SEAP as well as secreted luciferase:
monitor two promoters, or use one reporter as a normalization control

More Information

Applications

Homogeneous assay protocols
Time-course studies
Test multiple compounds
Suitable for downstream experiments with the same cells
Use the dual assay (which includes secreted alkaline phosphatase, as well as a secreted luciferase assay) to monitor two promoters
Use the dual assay to monitor one promoter with a built-in normalization control

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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Luciferase reporters—Ready-To-Glow secreted luciferase assay

Benefits of using a live-cell assay

Dual secreted reporter assay—luciferase & SEAP

Notice to purchaser

631781: Ready-To-Glow MetLuc Monoclonal Antibody

Notice to purchaser

Limit of detection for recombinant Metridia luciferase protein

High signal intensity and stability using secreted Metridia luciferase

Monitoring promoter activation using the sequence-optimized secreted Metridia luciferase reporter

Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System

Linear range of detection of Ready-To-Glow secreted Metridia luciferase from media supernatants

Use of secreted Metridia luciferase in a high-throughput screening (HTS) application

631740: Ready-To-Glow Automation Kit

Notice to purchaser

Similar secretion kinetics of Metridia secreted luciferase and SEAP enable accurate comparisons of the relative timing of promoter activity

Monitoring activation of two promoters simultaneously

631734: Ready-To-Glow Dual Secreted Reporter Assay

Notice to purchaser

Limit of detection for recombinant Metridia luciferase protein

High signal intensity and stability using secreted Metridia luciferase

Monitoring promoter activation using the sequence-optimized secreted Metridia luciferase reporter

Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System

Linear range of detection of Ready-To-Glow secreted Metridia luciferase from media supernatants

Use of secreted Metridia luciferase in a high-throughput screening (HTS) application

631728: Ready-To-Glow Secreted Luciferase Reporter Assay

Notice to purchaser

631727: Ready-To-Glow Secreted Luciferase Reporter Assay

Limit of detection for recombinant Metridia luciferase protein

High signal intensity and stability using secreted Metridia luciferase

Monitoring promoter activation using the sequence-optimized secreted Metridia luciferase reporter

Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System

Linear range of detection of Ready-To-Glow secreted Metridia luciferase from media supernatants

Use of secreted Metridia luciferase in a high-throughput screening (HTS) application

Notice to purchaser

Limit of detection for recombinant Metridia luciferase protein

High signal intensity and stability using secreted Metridia luciferase

Monitoring promoter activation using the sequence-optimized secreted Metridia luciferase reporter

Flow chart of the Ready-To-Glow Secreted Luciferase Reporter System

Linear range of detection of Ready-To-Glow secreted Metridia luciferase from media supernatants

Use of secreted Metridia luciferase in a high-throughput screening (HTS) application

631726: Ready-To-Glow Secreted Luciferase Reporter Assay

Overview

More Information

Applications

Additional product information