Ready-To-Glow Dual Secreted Reporter Assay
Ready-To-Glow Dual Secreted Reporter Assay
Brand: Takara Bio.
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Ready-To-Glow Dual Secreted Reporter Assay
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.
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
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
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.
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.
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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.
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.
