Dual Color ELISpot Kits

Assay Principle
View Full Assay Principle

R&D Systems Dual-Color ELISpot Kits are complete microplate-based immunoassays designed for the detection and enumeration of two different immune cell populations in a single sample. ELISpot assays are highly sensitive and can quantitate actively secreting cells even when cell frequencies fall below 1 in 100,000. Unlike other methods that assess antigen-specific immune cell responses, this assay does not require prior in vitro expansion of cells and is suitable for high-throughput analysis. These Dual-Color ELISpot Kits are complete ready-to-run assays and require no further development or refinement.

Features

  • Detect and quantitate individual cells secreting specific analytes
  • High sensitivity – ELISpot assays can measure responses with frequencies below 1 in 100,000 cells
  • No in vitro expansion of cells required
  • Amenable to high-throughput

R&D Systems Dual-Color ELISpot Kits employ antibody-coated, membrane-backed microplates to identify single cells that secrete either one or both analytes specified in the kit. The immobilized capture antibodies bind to cytokines that have been secreted in their vicinity, and enzyme conjugates are used to visualize the secreted proteins. Colored precipitates form at the sites of cytokine localization and appear as spots, with each individual spot representing an individual cytokine-secreting cell.

Dual Color Kit Contents

  • PVDF-backed 96-well Microplate
  • Detection Antibodies
  • Streptavidin-AP
  • BCIP/NBT
  • AEC
  • Positive Controls
  • Dilution Buffers
  • Wash Buffer

Detection of IFN-gamma and IL-17 Secretion by Mouse Splenocytes using the Dual-Color ELISpot Kit

Detection of IFN-gamma and IL-17 Secretion by Mouse Splenocytes using the Dual-Color ELISpot Kit IFN-gamma (blue spots) and IL-17 (red spots) were secreted from mouse splenocytes stimulated with PMA/Ca2+ ionomycin. Spots of cytokine secretion were visualized using the Mouse IFN-gamma/IL-17 Dual-Color ELISpot Kit (Catalog # ELD5007).