膜杰作
Star Staining
组分（Materials Provided）
| ID | Components | Size |
| EP169-C01 | High-bind Plate | 1 plate |
| EP169-C02 | Human IgE Fc | 60 μg |
| EP169-C03 | Human Fc epsilon RI alpha-SABC | 0.583 μg |
| EP169-C04 | Anti-IgE (Omalizumab) | 30 μg |
| EP169-C05 | Coating Buffer | 12 mL |
| EP169-C06 | 20×Washing Buffer | 50 mL |
| EP169-C07 | Blocking Buffer | 50 mL |
| EP169-C08 | Substrate Solution | 12 mL |
| EP169-C09 | Stop Solution | 7 mL |
应用说明（Application）
This kit is developed for screening for inhibitors of human IgE Fc binding to human Fc epsilon RI alpha.
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）
1. Unopened kit should be stored at 2℃-8℃ upon receiving.
2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
原理（Assay Principles）
High affinity immunoglobulin epsilon receptor subunit alpha (FCER1A) is also known as Fc-epsilon RI-alpha (FcERI), IgE Fc receptor subunit alpha, FCE1A. FCER1A contains two Ig-like (immunoglobulin-like) domains. FCER1A binds to the Fc region of immunoglobulins epsilon and is a high affinity receptor. FCER1A is responsible for initiating the allergic response, which binding of allergen to receptor-bound IgE leads to cell activation and the release of mediators (such as histamine) responsible for the manifestations of allergy. The same receptor also induces the secretion of important lymphokines. FCER1A plays a central role in allergic disease, coupling allergen and mast cell to initiate the inflammatory and immediate hypersensitivity responses that are characteristic of disorders such as hay fever and asthma.
Your experiment will include 4 simple steps:
1) Coat the plate with Human IgE Fc.
2) Add your molecule of interest to the tests.
3) Add Human Fc epsilon RI alpha-SABC to bind the coated Human IgE Fc.
4) Add TMB or other colorimetric HRP substrate.
Finally, the ability of your compound to inhibit IgE Fc : Fc epsilon RI alpha 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.
Serial dilutions of Anti-IgE (Omalizumab) (Catalog # EP169-C04) (1:1 serial dilution, from 5μg/mL to 0.078125μg/mL) was added into Human IgE Fc: Biotinylated Human Fc epsilon RI alpha 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).
For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The example data is for reference only.
背景（Background）：Fc epsilon RI alpha
High affinity immunoglobulin epsilon receptor subunit alpha (FCER1A) is also known as Fc-epsilon RI-alpha (FcERI), IgE Fc receptor subunit alpha, FCE1A. FCER1A contains two Ig-like (immunoglobulin-like) domains. FCER1A binds to the Fc region of immunoglobulins epsilon and is a high affinity receptor. FCER1A is responsible for initiating the allergic response, which binding of allergen to receptor-bound IgE leads to cell activation and the release of mediators (such as histamine) responsible for the manifestations of allergy. The same receptor also induces the secretion of important lymphokines. FCER1A plays a central role in allergic disease, coupling allergen and mast cell to initiate the inflammatory and immediate hypersensitivity responses that are characteristic of disorders such as hay fever and asthma.
