NovaBright™ Chemiluminescent Reporter Gene Assays

NovaBright™ Secreted Placental Alkaline Phosphatase (SEAP) kits contain the high-performance CSPD® alkaline phosphatase substrate, Emerald™ luminescence enhancer, and a unique buffer system that specifically inhibits endogenous nonplacental alkaline phosphatase activity. The persistent glow of the assay signal permits the use of simple luminometers without injectors.

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NEW! NovaBright™ Phospha-Light™ EXP

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

Performance

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.

SEAP Report Gene Assays - higher signal intensity
   
Figure 1. Higher signal intensity
 SEAP Report Gene Assays - higher signal/noise ratio
    
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.

Versatility

Use cells for other applications. Measure SEAP reporter enzyme in samples of culture medium removed from cells. Ideal for performing time course or multiparametric studies

-OR-



Measure SEAP reporter enzyme in the original culture well in the presence of cells, using the same assay protocol and reagent volumes (if cells are not being used for separate assay read-out)



Measure PLAP (non-secreted) reporter enzyme in cells

Assay demonstrated with numerous sample types (transiently transfected cell lines, primary cells, stably transfected cell lines, serum from transgenic animals) [4, 7, 12, 13, 15, 17, 18, 21, 22] See References

Wide range of applications—gene expression, promoter function, signal transduction pathway activation, gene knockdown, tissue-specific gene therapy targeting, viral function applications such as gene expression, cell-cell fusion, replication, antibody neutralization, infectivity, secretion pathway monitoring, quantitation of SEAP-tagged proteins [1, 8, 9, 10, 11, 14, 19, 20] See References

New Phospha-Light™ EXP Workflow vs. Original Phospha-Light

NovaBright™ Phospha-Light™ EXP Kit

NovaBright™ SEAP 2.0 Kit

(click to enlarge)

Original NovaBright™ Phospha-Light Kit
 

NovaBright™ Ordering Information

Sku Name Size Price Qty
T1015 Phospha-Light™ SEAP Reporter Gene Assay System 400 assays USD 262.00
T1016 Phospha-Light™ SEAP Reporter Gene Assay System 10,000 assays USD 3,875.00
T1017 Phospha-Light™ SEAP Reporter Gene Assay System 1,200 assays USD 574.00
N10577 NovaBright™ Phospha-Light™ EXP Assay Kit for SEAP (Secreted Placental Alkaline Phosphatase) Reporter Gene Detection 2 x 96 well plates USD 288.00
N10578 NovaBright™ Phospha-Light™ EXP Assay Kit for SEAP (Secreted Placental Alkaline Phosphatase) Reporter Gene Detection 10 x 96 well plates USD 1,124.00

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