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IL-13RA2 [Biotinylated] : IL-13 Inhibitor Screening ELISA Kit

For research use only.

IDComponentsSize
EP149-C01High-bind Plate1 plate
EP149-C02Human IL-13 55 μg
EP149-C03Biotinylated Human IL-13RA210 μg
EP149-C04Anti-IL-13RA2 Neutralizing Antibody20 μg
EP149-C05Streptavidin-HRP10 μg
EP149-C06Coating Buffer12 mL
EP149-C0710xWashing Buffer 50 mL
EP149-C08Blocking Buffer50 mL
EP149-C09Substrate Solution12 mL
EP149-C10Stop Solution7 mL

背景(Background)

Interleukin-13 receptor subunit alpha-2 is also known as IL13Rα2, IL13Ra2 cluster of differentiation 213A2, CD213A2, CT19, IL-13R, IL13BP, and is a membrane bound protein that in humans is encoded by the IL13RA2 gene. IL13Rα2 is closely related to IL13Rα1, a subunit of the interleukin-13 receptor complex. This protein binds IL13 with high affinity, but lacks any significant cytoplasmic domain, and does not appear to function as a signal mediator. It is, however able to regulate the effects of both IL13 and IL4, despite the fact it is unable to bind directly to the latter. It is also reported to play a role in the internalization of IL13. IL13Rα2 is a component of the cell surface receptors, however, the majority exists in intracellular pools and in soluble form, and thus plays an opposite role as a potent IL13 antagonist compared with IL13Rα1. It also functions as an inhibitor of IL4-dependent pathway probably through the physical interaction between the short intracellular domain of and cytoplasmic domain of IL13Rα2 and the IL4Rα chain. In spite of the failed STAT signaling function, IL13Rα2 dose induce TGF-beta production and fibrosis. Additionally, IL13Rα2has been reported to be abundantly and specifically overexpressed in glioblastoma multiforme.

应用说明(Application)

This kit is developed for screening for inhibitors of human IL-13 binding to human IL-13RA2.

It is for research use only.

重构方法(Reconstitution)

Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.

存储(Storage)

Unopened kit should be stored at 2°C-8°C upon receiving. Find the expiration date on the outside packaging and do not use reagents past their expiration date.

The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

原理(Assay Principles)

This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new IL-13 R alpha 2 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human IL-13 R alpha 2 to immobilized human IL-13 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated IL-13 R alpha 2 protein, a human IL-13 protein, an anti-IL-13 R alpha 2 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.

Your experiment will include 4 simple steps:

a) Coat the plate with human IL-13.

b) Add your molecule of interest to the tests.

c) Add human IL-13 R alpha 2-Biotin to bind the coated IL-13.

d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.

Finally, the half maximal inhibitory concentration (IC50) of your compound to IL-13 R alpha 2: IL-13 binding will be determined by comparing OD readings among different experimental groups.

 

典型数据-Typical Data Please refer to DS document for The assay protocol.

IL-13RA2 TYPICAL DATA

INHIBITION OF IL-13RA2[Biotinylated]: IL-13 BINDING BY ANTI-IL-13RA2 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-IL-13RA2 Neutralizing antibody (Catalog # EP149-C04) (1:1 serial dilution, from 10 μg/mL to 0.039μg/mL) was added into Biotinylated IL-13RA2:IL-13 binding reactions. The assay was performed according to the protocol described below. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).

 

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