The 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.
The Secrete Pair kit is compatible for use with GLuc-ON™ Promoter Reporter Clones
and miRNA Target clones
Figure 1. Comparison of GLuc signal stability in different buffer systems from Secrete-Pair and a competitor Gaussia luciferase assay kit. Cell culture medium was collected from cells transfected with humanized GLuc reporter clones. 10 µl of the medium was used in each assay. Two buffer systems of each kit were tested and the assays were performed according to the manufacturer protocols. The percentage of signal retained (Y axis) is used as an indicator for signal stability. For both kits, the GLuc activities in buffers with a stabilizer (-S) are much more stable than those in buffers without a stabilizer (-H). However, when compared side-by-side, Secrete-Pair buffer systems provide more stable GLuc signal (more than 90% of signal retained within the first 10 minutes) than the competitor kit kit.
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.