Invitrogen™ NovaBright™ Phospha-Light™ EXP Assay Kit for SEAP (Secreted Placental Alkaline Phosphatase) Reporter Gene Detection

Sensitive detection of alkaline phosphatase reporter enzyme enables a large number of applications in many areas of life science research, including gene expression, viral function assays, vaccine development, development of viral vectors and gene delivery methods for gene therapy, in vivo gene expression monitoring and novel cellular functional assays. This SEAP assay is one of the easiest and fastest methods for optimizing transfection efficiency. This assay has been used for a wide variety of viral functional assays, including viral gene expression assays, viral replication, viral fusogenicity, virus neutralization and viral-mediated cell-cell fusion, and viral infectivity. Use of the SEAP reporter protein is very enabling for in vivo reporter gene assays, by assaying serum samples from transgenic, transfected or viral vector-infected animals. The NovaBright reporter gene assay system has been used to measure SEAP levels in sera from transgenic or transfected whole animals, including mouse, rat, marmoset, monkey and pig sera, and in chicken egg allantoic fluid. The mouse SEAP protein (mSEAP) has recently been developed for improved SEAP protein stability in transgenic mice, and the NovaBright SEAP assay has been used for sensitive detection of mSEAP . In addition to reporter gene (gene expression) applications, the assay system is used to measure SEAP as a functional reporter for receptor-ligand binding assays with a SEAP-ligand chimera, protease-mediated secretion, and for secretion pathway activity. The NovaBright assay system can be used for the cellular measurement of non-placental alkaline phosphatase as a biomarker.

This kit provides a much simpler procedure than other assays. The assay is homogenous and requires only two solutions both of which are supplied ready-to-use. Prolonged glow light emission kinetics provides a stable light signal for read-time flexibility. The assay signal is measured in a simple luminometer without injectors. SEAP is a reporter protein that is secreted into the cell culture medium and detected by testing aliquots of the medium. Since SEAP is released into the medium, cell lysis is unnecessary. However, the assay also allows the detection placental alkaline phosphase (PLAP) isoenzyme following cell lysis.

• Second generation chemiluminescent reporter gene assay
• Simple, fast protocol, high throughput-compatible
• Ultrasensitive detection of secreted placental alkaline phosphatase
• Compatibility with automated platforms with trigger solution/positions already in place

• Detection Method: Chemiluminescent
• For Use With Equipment: Luminometer (Microplate)
• Format: 96-well plate
• High Throughput Compatibility: High Throughput-Compatible
• Readout: End Point
• Substrate: CSPD
• Substrate Properties: Chemical Substrate
• Substrate Type: AP (Alkaline Phosphatase) Substrate
• Target Enzyme: Placental Alkaline Phosphatase