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Cytokine Detection with Lumit™ Immunoassays

Lumit™ Immunoassays for cytokine detection quantitatively measure target analytes in cell culture samples with a simple, no-wash assay protocol. The assays can be performed directly on cell culture samples or on medium transferred from cell plates.

Lumit™ Cytokine Detection Immunoassay Overview

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

lumit-assay-principle

The protocol simply requires addition of the labeled antibodies to the sample, addition of the detection reagent and reading of the luminescent signal. The entire protocol is completed in less than 70 minutes.

lumit-assay-generic-workflow

Applications

lumit-il-1beta-thp-1-pbmcs

Lumit™ Immunoassay IL-1β detection in differentiated THP-1 cells and PBMCs. Panel A. THP-1 cells (ATCC; 5 x 104 cells/well) in a 96-well plate were differentiated with 20nM phorbol-12-myristate-13-acetate (PMA) for 3 days. Culture medium was replaced and various toll-like receptor (TLR) agonists were added overnight. IL-1β quantities were calculated using cubic spline interpolation. Panel B. PBMCs pooled from four donors (BioIVT) were plated at 6.5 x 104 cells/well in a 96-well plate and treated with various TLR agonists overnight. IL-1β quantities were calculated with cubic spline interpolation.

Benefits

  • 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.
  • Scalable and high throughput: Perform the assays in low volumes in 96-well or 384-well plates; precoated plates not required.
  • Luminescence detection: Use a standard plate-reading luminometer; no specialized instruments or modules required.

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

E-mail Promega

Available Cytokine Detection Immunoassays

The following assays are in development and may be available as early-access materials or by custom order.

  • IL-1β (human) immunoassay
  • IL-1β (mouse) immunoassay
  • IL-2 (human) immunoassay
  • IL-6 (human) immunoassay
  • IFN-γ (human) immunoassay
  • TNF-α (human) immunoassay
  • IL-4 (human) immunoassay
  • IL-10 (human) immunoassay
  • Please contact us for more information about these assays and to learn about availability.

    E-mail Promega