NovaBright Chemiluminescent Secreted Placental Alkaline Phosphatase (SEAP) Reporter Gene Assays

SEAP Phospha-Light EXP and Original SEAP Phospha-Light Kit Benefits:

• Designed to detect SEAP reporter gene expression
• Measurement of either sample culture media or the total culture (cell and media)
• No cell lysis steps—your samples remain intact
• Easy and fast assay: <30 minute assay time, <1.5 hours to results
• Wide linear dynamic range provides 3 orders of magnitude greater sensitivity than colorimetric methods
• Proprietary buffer ensures low background—no signal from endogenous alkaline phosphatases
• Persistent glow signal lasts for easy plate reading—no injectors required, assay is easily scaled up for higher-throughput plate formats
• Compatible with Phenol red

New SEAP Phospha-Light EXP Assay Format Includes:

• Ready-to-use CSPD substrate and next-generation Emerald enhancer
• Only two assay reagents for simplicity
• No dilution step
• Significantly shorter heat-inactivation step

The new NovaBright Phospha-Light EXP kit provides higher signal intensity (Figure 1), higher signal/noise ratio, and low-end sensitivity compared to the original NovaBright Phospha-Light kit and other commercially available luminescent SEAP assay kits.

Figure 1. Higher signal intensity		Figure 2. Provides higher signal/noise ratio and low-end sensitivity.
Figure 3. The SEAP reporter gene allows for time-course and/or assay development studies that are inconvenient using assays that depend on cell destruction. Here, dose response of NFkB signaling in response to treatment with TNFa was monitored at different time points. HEK293 cells were transfected with a pNFkB-SEAP construct in a 24-well tissue-culture dish. At 48 hrs post transfection, media was removed and replaced with TNFa-containing media.  Culture media was removed and assayed for SEAP enzyme activity using the NovaBright Phospha-Light–EXP Assay at the indicated time points.		Figure 4. Because SEAP reporter gene expression is monitored using cell culture media or supernatant, the cells can be used for other experiments.  Here we show sequential monitoring of forskolin-induced gene expression using the Phospha-Light-EXP SEAP assay to monitor SEAP activity in culture media and cell viability using the Invitrogen PrestoBlue assay on the cells remaining in the culture well. NIH/3T3 cells were transfected with pCRE-SEAP for cAMP-inducible SEAP expression using Lipofectamine LTX reagent with PLUS reagent.  Cells were treated with forskolin for 24 hours to induce cAMP production.

NovaBright™ Phospha-Light™ EXP Kit

Original NovaBright™ Phospha-Light™ Kit