Mouse CD4+/IL-4 ELISpot Kit

Name: Mouse CD4+/IL-4 ELISpot Kit
Brand: R&D Systems
SKU: EL6060

R&D Systems | Catalog # EL6060

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Product Documents

Discontinued Product

EL6060 has been discontinued. View all IL-4 products.

Product Summary
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species

Mouse

Assay Type

Quantitative Sandwich ELISA

Format

96-well PVDF-backed microplate

Assay Length

3 hours 15 mins to 4 hours 30 mins (plus plate coating)

Sample Type

Whole Cells

Product Summary for Mouse CD4+/IL-4 ELISpot Kit

PRODUCT SUMMARY
ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. This kit is designed for the detection and enumeration of mouse IL-4. Complete ELISpot kits are ready-to-run and require no assay development or refinement.

This ELISpot assay employs a capture antibody specific for mouse IL-4, pre-coated onto a PVDF-backed microplate. Appropriately stimulated cells are pipetted directly into the wells and the immobilized antibody in the immediate vicinity of the secreting cells binds secreted mouse IL-4. Following wash steps and incubation with a biotinylated detection antibody, alkaline-phosphatase conjugated to streptavidin is added. Unbound enzyme is subsequently removed by washing and a substrate solution (BCIP/NBT) is added. A blue-black colored precipitate forms at the sites of cytokine localization and appears as spots, with each individual spot representing an individual mouse IL-4 secreting cell. The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.

PRODUCT FEATURES

Detect and quantitate individual cells secreting mouse IL-4
High sensitivity - ELISpot assays can measure responses with frequencies well below 1 in 100,000 cells
No in vitro expansion of cells required
High-throughput - ELISpot assays use only a small number of primary cells

KIT CONTENTS

Mouse IL-4 Microplate
Biotinylated Detection Antibody
Streptavidin conjugated to Alkaline Phosphatase
Dilution Buffers
Wash Buffer Concentrate
BCIP/NBT Chromogen
Mouse IL-4 Positive Control

OTHER REAGENTS REQUIRED

Pipettes and pipette tips
Deionized or distilled water
Squirt bottle, manifold dispenser, or automated microplate washer
500 mL graduated cylinder
37 °C CO₂ incubator
Sterile culture media
Dissection microscope or an automated ELISpot reader

Formulation, Preparation, and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-4

IL-4 (Interleukin-4) is a cytokine that is primarily expressed by Th2-biased CD4⁺ T cells, mast cells, basophils, and eosinophils. It promotes cell proliferation, survival, and immunoglobulin class switch to IgG4 and IgE in human B cells, acquisition of the Th2 phenotype by naÃ¯ve CD4⁺ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells. IL-4 plays a dominant role in the development of allergic inflammation and asthma. The type I receptor for IL-4, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 R alpha and the common gamma chain (a shared subunit of the receptors for IL-2, -7, -9, -15, and -21). The type II receptor on nonhematopoietic cells consists of IL-4 R alpha and IL-13 R alpha 1. The type II receptor also transduces IL-13 mediated signals.