Porcine IL-10 ELISpot Kit

Catalog #: EL693 Datasheet / COA / SDS

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EL693

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Porcine IL-10 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 porcine IL-10. Complete ELISpot kits are ready-to-run and require no assay development or refinement.

This ELISpot assay employs a capture antibody specific for porcine IL-10, 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 porcine IL-10. 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 porcine IL-10 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 porcine IL-10
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

Porcine IL-10 Microplate
Biotinylated Detection Antibody
Streptavidin conjugated to Alkaline Phosphatase
Dilution Buffers
Wash Buffer Concentrate
BCIP/NBT Chromogen
Porcine IL-10 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-10

Interleukin-10 functions as an anti-inflammatory cytokine by inhibiting the expansion and activation of Th1 cells and Th17 cells and by promoting the development of M2 macrophages and regulatory T cells (Treg). Within a tumor microrenvironment, however, IL-10 can inhibit the expansion of both Treg and myeloid-derived suppressor cells (MDSC). IL-10 exerts protective effects including limiting tissue damage in arthritic inflammation and promoting muscle regeneration after injury, but it also contributes to the persistence of viral infections. IL-10 signals through a receptor complex composed of IL-10 R alpha and IL-10 R beta. IL-10 R beta additionally associates with IL-20 R alpha, IL-22 R alpha 1, or IL-28 R alpha to form the receptor complexes for IL-22, IL-26, IL-28, and IL-29.

Long Name:

Interleukin 10

Entrez Gene IDs:

3586 (Human); 16153 (Mouse); 25325 (Rat); 397106 (Porcine); 403628 (Canine); 102133450 (Cynomolgus Monkey); 493683 (Feline); 100715618 (Guinea Pig); 2949786 (Viral)

Alternate Names:

CSIF; CSIFMGC126450; Cytokine synthesis inhibitory factor; GVHDS; IL10; IL-10; IL10A; IL-10MGC126451; interleukin 10; interleukin-10; TGIF