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Detection of Metabolic Regulators with Lumit™ Immunoassays

The Lumit™ Insulin and Glucagon Immunoassays are homogeneous, bioluminescent ELISA alternatives to measure insulin and glucagon from pancreatic islets and cells in culture. The assays offer a fast, easy, no-wash protocol. Simply add Lumit™ reagents to your sample, incubate and read using a standard multimode plate reader. Results are available in just over an hour, so you can plan your next experiment sooner. Lumit™ assays are ideal for quantitating hormone secretion from islet perifusion workflows. The flexible assay format can be scaled to 384-well plates for high-throughput sample processing.

Product Availability

The Lumit™ Glucagon Immunoassay is available through our catalog.

Learn More or Request a Sample

The Lumit™ Insulin Immunoassay is available as early access material. Contact us for a current list of available kits and ordering information.

Enquire About Product and Sample Availability

How the Lumit™ Immunoassay Works

Primary antibodies to the analyte, selected for their specificity and sensitivity, 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. Adding the optimized luciferase substrate generates a bright luminescent signal that is proportional to analyte levels. The entire immunoassay is completed in just 70 minutes.

lumit-assay-principle
Lumit assay workflow.

In this publication, Tong and Stein use the Lumit™ Insulin Immunoassay to investigate of the impact of lipid droplet formation on pancreatic islet cells.

Tong, X. and Stein, R. (2021) Lipid droplets protect human β-cells from lipotoxicity-induced stress and cell identity changes. Diabetes 70, 2595–607.

See the Data

In this work, El and colleagues used the Lumit™ Glucagon Immunoassay to characterize the complex regulatory mechanisms that control glucagon secretion from alpha cells.

El, K. et al. (2021) GIP mediates the incretin effect and glucose tolerance by dual actions on α-cells and β-cells. Sci. Adv. 7, abf1948.

See the Data Read More About this Work

Benefits

  • Scalable and high throughput: Perform the assays in low volumes in 96-well or 384-well plates; precoated plates not required.
  • No wash: Simple add-mix-read protocol.
  • 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 most cell models.
  • Luminescence detection: Use a standard plate-reading luminometer; no specialized instruments or modules required.

Using the Lumit™ Immunoassay to Detect Hormone Secretion

Insulin and glucagon are key target analytes measured in metabolism research, including Diabetes and Liver Disease Research. Secreted from β- and α-cells of pancreatic islets, respectively, the two hormones are vital for regulating glucose levels in the body.

Using the Lumit™ Immunoassays, it is easy to collect hormone secretion data from monolayer cells in culture, 3D islet microtissues and islets. The simple add-and-read protocol is compatible with static and dynamic glucose-stimulated insulin secretion (GSIS) experiments, including perifusion systems.

Quantitate Insulin Secretion from INS-1 Cells

Bar graph showing glucose stimulated insulin secretion in INS-1 cells with increasing concentrations of glucose and in varying cells densities.
Bar graph showing increasing insulin release in INS-1 cells with increasing glucose concentration.

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 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.

Quantitate Insulin and Glucagon Secretion from Mouse Pancreatic Islets

Representative perifusion data showing insulin and glucagon secretion from mouse islets.

Insulin and glucagon secretion were measured in samples collected during perifusion experiments. Briefly, 80 mouse islets were placed in triplicate chambers of a perifusion instrument (Biorep, Miami Lakes, Florida). The islets were treated with 2.7mM glucose and then 10mM glucose, in combination with an amino acid mixture. Perifusate was collected every minute. Ten microliters of each sample were transferred into wells of a 384-well plate and assayed for either insulin or glucagon. Insulin and glucagon data sets are superimposed. This data was kindly provided by Drs. Hannah Foster and Matthew Merrins, University of Wisconsin VA Hospital, Madison, Wisconsin.