ID | Components | Size |
EP159-C01 | High-bind Plate | 1 plate |
EP159-C02 | Human IL-11 | 55 μg |
EP159-C03 | Biotinylated Human IL-11R | 20 μg |
EP159-C04 | Anti-IL-11 Neutralizing Antibody | 20 μg |
EP159-C05 | Streptavidin-HRP | 5 μg |
EP159-C06 | Coating Buffer | 12 mL |
EP159-C07 | 10xWashing Buffer | 50 mL |
EP159-C08 | Blocking Buffer | 50 mL |
EP159-C09 | Substrate Solution | 12 mL |
EP159-C10 | Stop Solution | 7 mL |
背景(Background)
nterleukin-11 (IL-11) is a pleiotropic cytokine that stimulates megakaryocytopoiesis, resulting in increased production of platelets, as well as activating osteoclasts, inhibiting epithelial cell proliferation and apoptosis, and inhibiting macrophage mediator production. These functions may be particularly important in mediating the hematopoietic, osseous and mucosal protective effects of IL-11. The cytokine also possesses anti-inflammatory activity, and has been proposed as a therapeutic agent in the treatment of chronic inflammatory diseases, such as Crohns disease and rheumatoid arthritis. Although IL-11 was initially believed to be restricted to mammals, subsequent studies demonstrated it to be expressed in fish. Despite close similarity in gene structure and conservation of key amino acids between fish and mammalian IL-11, they share relatively low overall amino acid identity and may not necessarily be functionally analogous.
应用说明(Application)
This kit is developed for screening for inhibitors of human IL-11 binding to human IL-11R.
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-11 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human IL-11R to immobilized human IL-11 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated IL-11R protein, a human IL-11 protein, an anti-IL-11 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human IL-11.
b) Add your molecule of interest to the tests.
c) Add human IL-11R-Biotin to bind the coated IL-11.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to IL-11:IL-11R 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.
INHIBITION OF IL-11: IL-11R [Biotinylated] BINDING BY ANTI-IL-11 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-IL-11 Neutralizing antibody (Catalog # EP159-C04) (1:1 serial dilution, from 10 μg/mL to 0.0195μg/mL) was added into IL-11: Biotinylated IL-11R 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).