cAMP Gs dynamic kit
Optimal performance when applied to Gs activation or inhibition
Active GLP-1, also known as glucagon-like peptide-1 active forms (GLP-1(7-36)NH2, and GLP-1(7-37)), has become a key biomarker in treatment of type 2 diabetes. The main actions of GLP-1 are stimulation of insulin secretion, and inhibition of glucagon secretion and food intake. In vivo, the active forms are rapidly degraded into inactive forms (9-36)NH2 and (9-37) by the dipeptidyl peptidase IV (DPP-IV). The active GLP-1 kit is designed for a rapid detection of GLP-1 in cell supernatants.
|Incubation||ON at RT|
|Linear range||43. 8 to 1,400 pg/mL|
|Detection limit||25 pg/mL*|
|Species reactivity||Rat, mouse, human, porcine|
|GLP-1 (7-36) amide||100%|
|GLP-1 (1-36) amide||0.39%|
|GLP-1 (9-36) amide||<0.01%|
Active GLP-1 standard curve was performed in a final 20µL assay volume and read on a Pherastar instrument (BMG LABTECH).
* Caution: all HTRF compatible readers but SpectraMax M5e and FlexStation reach this sensitivity criteria.
|HTRF active GLP-1||Fluorescent ELISA|
|1 assay step mix and read (no washing) - <1hrs assay time||6 sequential steps - >4hrs assay time|
A time-course was determined for active GLP-1 secretion in human enteroendocrine NCI-H716 cells after glucose stimulation. Cells (2.105 cells/well) were seeded into 96-well culture plates pre-coated with Matrigel, and cultured for two days with DMEM 10% FBS medium. On the experiment day, cells were washed and incubated for 2 hours with increasing concentration of glucose in 100 µL of KRB supplemented with DPP IV inhibitor . At the end of glucose stimulation, 10 µL of supernatants were transferred to a 384 small volume-well plate, and assayed for active GLP-1 level as described in 'Assay protocol'. As expected, active GLP-1 was induced with Glucose dose dependence. The maximum amplitude of active GLP-1 triggered by 10% glucose versus basal production was 2.1.
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