膜杰作
Star Staining
| ID | Components | Size |
| EP115-C01 | Pre-coated Human ACE2 Microplate | 1 plate |
| EP115-C02 | SARS-CoV-2 Inhibitor | 20 μg |
| EP115-C03 | HRP-SARS-CoV-2 Spike RBD(B.1.1.529) | 15 μg |
| EP115-C04 | 10xWashing Buffer | 50 mL |
| EP115-C05 | Dilution Buffer | 50 mL |
| EP115-C06 | Substrate Solution | 12 mL |
| EP115-C07 | Stop Solution | 7 mL |
背景（Background）
As of Nov 29th, confirmed cases of a new variant of concern (VOC) B.1.1.529 have been reported in South Africa, Botswana and more than 12 other countries. Thanks to the rapid sequencing efforts of South African scientists, the whole world is now alerted about this potentially devastating lineage, which was assigned the name Omicron by WHO.Omicron is a highly divergent variant with a high number of mutations (26-35) in the spike. Identification of some familiar mutations that also appeared on other VOCs gives rises to concerns of its immune escape potential and higher transmissibility,it is necessary to develop SARS-CoV-2 inhibitors, such as vaccines，therapeutic antibodies and small molecule compounds against SARS-CoV-2(B.1.1.529). The SARS-CoV-2 (B.1.1.529) Inhibitor Screening Kit (Spike RBD) is helpful to develop the to test SARS-CoV-2 inhibitors.
应用说明（Application）
This kit is developed for screening inhibitors of SARS-CoV-2 (B.1.1.529).
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 & 运输（Storage & Shipping）
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）
This assay kit is used to measure the levels of Anti-SARS-CoV-2 neutralizing antibody through a competitive ELISA. The microplate in the kit has been pre-coated with Human ACE2 protein. First samples are added to the wells followed by addition of HRP-SARS-CoV-2 Spike RBD. After incubation, the wells are washed and substrate is added to the wells. The reaction is terminated by the addition of stop solution and the intensity of color is measured at 450 nm. The presence of neutralizing antibodies in samples will compete with ACE2 for HRP-SARS-CoV-2 Spike RBD binding. The intensity of assay signal decrease proportionally to the presence of Anti-SARS-CoV-2 neutralizing antibody.
Your experiment will include 5 simple steps:
a) All reagents were returned to room temperature(20°C-25°C) before use.
b) Make series the tested sample and SARS-CoV-2 Inhibitor with ditlution buffer, HRP-SARS-CoV-2 RBD diluted with dilution buffer.
c) Add the diluted sample, Control and the HRP-SARS-CoV-2 RBD add the plate respectively.
d) Wash the plate and add TMB or other colorimetric HRP substrate.
e) Stop the substrate reaction by add diluted acid. Absorbance (OD) is calculated as the absorbance at 450 nm minus the absorbance at 630 nm to remove background prior to statistical analysis. The OD Value reflects the amount of antibody bound.
典型数据-Typical Data Please refer to Ds document for the assay protocol.
Incubation of SARS-CoV-2 inhibitor:Blocking the binding of HRP-SARS-CoV-2 Spike RBD(B.1.529) to Human ACE2.
Serial dilutions of SARS-CoV-2 inhibitor (Catalog # EP115-C02) (1:1 serial dilutions, from 25ug/mL to 0.049ug/mL(171.9nM to 0.34nM)) and loaded onto the plate coated by human ACE2 in the presence of HRP-SARS-CoV-2 Spike RBD(B.1.1.529). Background was subtracted from data points prior to log transformation and curve fitting.
