Takara Great EscAPe SEAP Chemiluminescence Kit 2.0 \1,000 Rxns

The Great EscAPe SEAP Chemiluminescence Kit 2.0 is a complete system for the detection of secreted alkaline phosphatase (SEAP) activity in the growth media from mammalian cells expressing this enzyme. The kit contains reagents necessary for the chemiluminescent detection of SEAP activity using a 96-well plate luminometer. The assay can also be performed in single tubes and measured in a tube luminometer. It allows you to monitor promoter activity over time, because samples of the cell media can be taken repeatedly and tested without lysing cells.

SEAP activity is highly specific, with a >20-fold increase in activity.HEK 293 cells were transiently transfected with a promoter construct containing CRE driving the expression of SEAP, or mock-transfected. 12 hr posttransfection, the cell culture media was replaced by media containing either 10 µm forskolin or plain cell culture media and incubated for 7 hr. Forskolin causes an increase in the level of cytosolic cAMP, which in turn activates CRE, driving the expression of SEAP, which is detectable in the media culture supernatant. The samples were assayed using the Great EscAPe SEAP Chemiluminescence Kit 2.0 and its protocol and analyzed on a BD Monolight 3096 Luminometer.

Comparison of the sensitivities of SEAP and firefly luciferase.Parallel cultures of BHK cells were transiently transfected with the indicated amounts of either pSEAP2-Control Vector or a similar vector expressing luciferase. After 24 hr, SEAP activity was assayed in the appropriate culture media using the Great EscAPe chemiluminescent assay. Similarly, after 24 hr, cell lysates were prepared from the luciferase cultures, and luciferase activity was assayed using a commercial kit. RLU = relative light units.