| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| ACE2 Protein Coated Microwells | 30924 | 96 tests |
|
+4C |
| SARS-CoV-2 Spike RBD Protein, HRP-linked | 42530 | 1 ea |
|
+4C |
| Sample Diluent A | 71637 | 25 ml |
|
+4C |
| HRP Diluent | 13515 | 11 ml |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Kit #69486 Positive Control | 70532 | 1 ea |
|
+4C |
| ELISA Kit #69486 Negative Control | 81889 | 1 ea |
|
+4C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit allows for the detection of antibodies (or small molecules) that block the interaction between the host receptor-binding domain (RBD) of the SARS-CoV-2 spike protein and ACE2. In this assay, SARS-CoV-2 spike RBD protein linked to HRP (RBD-HRP) is pre-incubated with the sample and controls, allowing blocking antibodies present in the sample to bind to the RBD-HRP. These pre-incubated mixtures are then added to microwell plates coated with ACE2 protein. The RBD-HRP is captured by ACE2 in the well to varying degrees depending on the blocking activity present in each sample. The wells are then washed to remove unbound material. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is inversely proportional to the ability of the sample to block the interaction between SARS-CoV-2 spike RBD and ACE2 proteins. For example, if no blocking activity is present in the sample, the RBD-HRP will be able to bind to ACE2 on the microwell plates, resulting in high signal. Conversely, samples with high blocking activity will prevent the RBD-HRP from binding to ACE2 on the microwell plates, resulting in low signal. This kit is designed to detect blocking antibodies present in human serum/plasma samples. However, this kit may also be used to assess the blocking activity of non-human antibodies and small molecules. In the latter scenarios (for non-human antibodies and small molecules), the user will have to determine the appropriate dilution/concentration of their samples to use, along with running the proper controls.
Specificity/Sensitivity
Background
The cause of the COVID-19 pandemic is a novel and highly pathogenic coronavirus, termed SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2). SARS-CoV-2 is a member of the Coronaviridae family of viruses (1). The genome of SARS-CoV-2 is similar to other coronaviruses, and is comprised of four key structural proteins: S, the spike protein, E, the envelope protein, M, the membrane protein, and N, the nucleocapsid protein (2). Coronavirus spike proteins are class I fusion proteins and harbor an ectodomain, a transmembrane domain, and an intracellular tail (3,4). The highly glycosylated ectodomain projects from the viral envelope surface and facilitates attachment and fusion with the host cell plasma membrane. The ectodomain can be further subdivided into host receptor-binding domain (RBD) (S1) and membrane-fusion (S2) subunits, which are produced upon proteolysis by host proteases at S1/S2 and S2’ sites. S1 and S2 subunits remain associated after cleavage and assemble into crown-like homotrimers (2,4). In humans, both SARS-CoV and SARS-CoV-2 spike proteins utilize the angiotensin-converting enzyme 2 (ACE2) protein as a receptor for cellular entry (5-7). Spike protein subunits represent a key antigenic feature of coronavirus virions, and therefore represent an important target of vaccines, novel therapeutic antibodies, and small-molecule inhibitors (8,9).
- Zhou, P. et al. (2020) Nature 579, 270-3.
- Tortorici, M.A. and Veesler, D. (2019) Adv Virus Res 105, 93-116.
- Li, F. et al. (2006) J Virol 80, 6794-800.
- Li, F. (2016) Annu Rev Virol 3, 237-61.
- Shang, J. et al. (2020) Nature 581, 221-4.
- Wrapp, D. et al. (2020) Science 367, 1260-3.
- Yan, R. et al. (2020) Science 367, 1444-8.
- Yuan, Y. et al. (2017) Nat Commun 8, 15092.
- Amanat, F. and Krammer, F. (2020) Immunity 52, 583-9.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
限制使用
除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。
专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专
