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Detection of Metabolic Regulators with Lumit™ Immunoassays

Insulin and glucagon are key target analytes measured in metabolism research. Secreted from β and α cells of pancreatic islets, respectively, the two hormones are vital for regulating glucose levels in the body. The Lumit™ Immunoassays for metabolic regulators quantitatively measure insulin and glucagon in cell culture samples with a scalable, no-wash assay protocol. These assays are ideally suited to process samples from glucose-stimulated insulin secretion and glucagon secretion experiments, including high-throughput perifusion assays.

Lumit™ Metabolic Regulator Immunoassay Overview

Primary antibodies, selected for their specificity and sensitivity, are used to detect the analyte of interest. The antibodies are labeled with the LgBiT and SmBiT subunits of NanoBiT® Luciferase. In the presence of the analyte, the subunits are brought together to reconstitute active luciferase enzyme. Upon addition of the optimized luciferase substrate, a bright luminescent signal is generated.

lumit-assay-principle

Lumit™ Immunoassays are homogeneous, no-wash assays. The protocol simply requires addition of the labeled antibodies to the sample, addition of the detection reagent (which includes the luciferase substrate) and then reading of the luminescent signal. The entire immunoassay is completed in less than 70 minutes.

lumit-assay-generic-workflow2

Lumit™ Assays to Detect Metabolic Regulators

Analytes available:

  • Lumit™ Insulin Immunoassay
  • Lumit™ Glucagon Immunoassay

Please contact us for more information about these assays and to learn about availability.

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Applications: Lumit™ Immunoassay to Detect Insulin Secretion

Using our Lumit™ Immunoassay, it is easy to collect hormone secretion data from monolayer cells in culture (data shown below), 3D islet microtissues and isolated islets. The simple add-and-read protocol is compatible with low- and high-throughput glucose-stimulated insulin secretion (GSIS) experiments, including perifusion systems.

lumit-insulin-assay-ins1
lumit-insulin-assay-glucose

Monitoring cellular insulin secretion response to glucose with Lumit™ Immunoassays. Panel A. INS-1 cells plated in duplicate at varying cells/well in 96-well plates were used for a glucose-stimulated insulin secretion experiment. After 60 minutes, 10µl of supernatant was removed and assayed with the Lumit™ Insulin Immunoassay Kit in 384-well plates. Error bars are +/–1 s.d. Panel B. The data from the same experiment using 10,000 cells/well is shown, demonstrating increasing release of insulin with increasing concentration of glucose.

Benefits

  • Scalable and high throughput: Perform the assays in low volumes in 96-well or 384-well plates; precoated plates not required.
  • No wash, no transfer: The simple add-mix-read protocol can be performed directly on cell cultures.
  • Rapid time to results: Get results just 70 minutes after starting the assay.
  • Large linear range: The picomolar to nanomolar linear range covers expected concentrations for the majority of cell models.
  • Luminescence detection: Use a standard plate-reading luminometer; no specialized instruments or modules required.

Lumit™ Immunoassays for metabolic regulators are available as early access material. Contact us for a current list of available kits and ordering information.

E-mail Promega