Secrete-Pair™ Dual Luminescence Assay Kit is designed to analyze the activities of Gaussia Luciferase (GLuc) and Secreted Alkaline Phosphatase (SEAP) of a dual-reporter system side-by-side using the same sample from the cell culture medium. Both GLuc and SEAP are secreted reporter proteins. Samples can be easily obtained from cell culture medium without lysis of the cells.
Two buffer conditions are provided in the kit for GLuc assays depending on the applications. Buffer GL-S contains a stabilizer and can be used for stabilized activity by overcoming the quick decay of the GLuc signal. When higher sensitivity is required for detecting low expression of GLuc, Buffer GL-H can be used for higher enzyme activity.
Secrete-Pair measures dual reporter signals and allows transfection normalization. The normalized GLuc activities can be compared across samples free of the impact of transfection variation.
Secrete Pair™ Gaussia Luciferase Assay Kit
The Secrete Pair Gaussia Luciferase Assay Kit is available for Gaussia luciferase assay alone as well.
The Secrete Pair kits are compatible for use with GLuc-ON™ promoter reporter clones and miTarget™ miRNA 3' UTR target clones.
Figure 2. GLuc and SEAP assays.
Cell culture medium was collected from cells transfected with GLuc-SEAP dual-reporter clone. 10 µl of the medium was used in each assay. At the beginning, the wtGLuc activity in Buffer GL-H is about 3-5 times higher than that in Buffer GL-S. Then it quickly decays. The GLuc activity in Buffer GL-S, however, is much more stable. The amount of SEAP substrate was adjusted so that the reading of SEAP and that of wtGLuc (in buffer GL-S) are at similar levels for normalization purpose.
