Canine IL-4 ELISpot Kit

Catalog #: EL754
1 Citation Datasheet / COA / SDS

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Summary Product Datasheets Preparation and Storage Background

Canine IL-4 ELISpot Kit Summary

Assay Type

Quantitative Sandwich ELISA

Format

96-well PVDF-backed microplate

Assay Length

3 hours 15 mins to 4 hours 30 mins*

Sample Type

Whole Cells

Specificity

Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

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 canine 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 canine 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 canine 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 canine 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 canine 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

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

Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Shipping

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

Stability & 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 naïve 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? and the common ? 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? and IL-13 R?1. The type II receptor also transduces IL-13 mediated signals.

Long Name:

Interleukin 4

Entrez Gene IDs:

3565 (Human); 16189 (Mouse); 287287 (Rat); 397225 (Porcine); 280824 (Bovine); 403785 (Canine); 574281 (Primate); 100302454 (Rabbit)

Alternate Names:

B cell growth factor 1; BCDF; B-cell stimulatory factor 1; BCGF1; BCGF-1; binetrakin; BSF1; BSF-1; IL4; IL-4; IL-4B_cell stimulatory factor 1; IL4E12; interleukin 4; interleukin-4; Lymphocyte stimulatory factor 1; MGC79402; pitrakinra

⚠ WARNING: This product can expose you to chemicals including methanol, which is known to the State of California to cause reproductive toxicity with developmental effects. For more information, go to www.P65Warnings.ca.gov.

Citation for Canine IL-4 ELISpot Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

Long-term correction of inhibitor-prone hemophilia B dogs treated with liver-directed AAV2-mediated factor IX gene therapy.
Authors: Niemeyer GP, Herzog RW, Mount J, Arruda VR, Tillson DM, Hathcock J, van Ginkel FW, High KA, Lothrop CD
Blood, 2008-10-28;113(4):797-806.
Species: Canine
Sample Types: Whole Cells