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Secreted Alkaline Phosphatase Assay

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The Great EscAPe SEAP 2.0 System uses a secreted alkaline phosphatase assay to analyze cis-acting DNA sequences and trans-acting factors. The kinetics of gene expression can easily be studied with this secreted alkaline phosphatase assay by repeatedly sampling the same culture. The kit has an extremely low background signal, with a wide dynamic range and high sensitivity. Furthermore, the robust signal is extremely stable over time. Cells can be studied further after the secreted alkaline phosphatase assay using Northern blots, RNase protection assays, Western blots and other methods.

 

Fluorescence-Based Detection

The Great EscAPe SEAP Fluorescence Detection Kit uses the fluorescent substrate 4-methylumbelliferyl phosphate (MUP) and has sensitivity comparable to firefly luciferase assays. Like the chemiluminescence assay, the secreted alkaline phosphatase fluorescence assay is linear over a 104-fold range of enzyme concentrations.

Dual Secreted Reporter Assay—SEAP & Luciferase

The Ready-To-Glow Dual Secreted Reporter System has two live cell reporters that do not require cell lysis. In addition to secreted alkaline phosphatase, it includes Metridia luciferase, which has a 2–4 fold higher signal compared to Renilla and firefly luciferases.


Cat. #  Product  Contents Size License
631736 Great EscAPe™ SEAP Chemiluminescence Kit 2.0 50 Rxns
631737 Great EscAPe™ SEAP Chemiluminescence Kit 2.0 300 Rxns
631738 Great EscAPe™ SEAP Chemiluminescence Kit 2.0 1,000 Rxns
631704 Great EscAPe™ SEAP Fluorescence Detection Kit 300 Rxns
631734 Ready-To-Glow™ Dual Secreted Reporter Assay 500 Rxns
631735 Ready-To-Glow™ Dual Secreted Reporter Vector Kit 4 x 20 ug
631715 pSEAP2-Basic Vector 20 ug
631717 pSEAP2-Control Vector 20 ug
Characterization:

Features

  • One step assay, for monitoring promoter activity
  • 10-fold higher sensitivity then firefly luciferase
  • No-cell-lysis protocol
  • Adaptable from single-tube to high-throughput assays
  • Dual, live cell assay includes secreted luciferase as well as SEAP:
    monitor two promoters, or use one reporter as a normalization control

Applications

  • Time course studies
  • Non-invasive reporter gene assays with secreted alkaline phosphatase
  • Test multiple compounds
  • Perform downstream experiments with the same cells
  • Use the dual system (which includes secreted luciferase, as well a secreted alkaline phosphatase assay) to monitor two promoters
  • Use the dual system to monitor one promoter with a built-in normalization control


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