Revision 15

#69486

Store at +4C

SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
All

UniProt ID:
#P0DTC2

Entrez-Gene Id:
#43740568

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Color Storage Temp
ACE2 Protein Coated Microwells3092496 tests+4C
SARS-CoV-2 Spike RBD Protein, HRP-linked425301 eaRed (Lyophilized)+4C
Sample Diluent A7163725 ml+4C
HRP Diluent1351511 mlRed+4C
ELISA Wash Buffer (20X)980125 ml+4C
TMB Substrate700411 ml+4C
STOP Solution700211 ml+4C
Sealing Tape545032 ea+4C
ELISA Kit #69486 Positive Control705321 ea+4C
ELISA Kit #69486 Negative Control818891 ea+4C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

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

The SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit detects endogenous levels of antibodies that block the interaction between the SARS-CoV-2 spike RBD (318-541) protein and human ACE2 (18-615). This kit is designed to detect blocking antibodies present in human serum/plasma, however, it may also be used to assess blocking activity of non-human antibodies and small molecules.

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).

Background References

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.