|1 Kit (96 assays)
|Spike Protein Coated Microwells II
|Anti-Human IgM, HRP-linked Antibody (ELISA Formulated)
|Sample Diluent A 71637
|ELISA Wash Buffer (20X) 9801
|TMB Substrate 7004
|STOP Solution 7002
|ELISA Kit #37322 Positive Control
|ELISA Kit #37322 Negative Control
NOTE: Prepare solutions with deionized/purified water or equivalent. Prepare only as much reagent as needed on the day of the experiment.
NOTE: Equilibrate all materials and prepared reagents to room temperature prior to running the assay.
Human-sourced samples should be handled in accordance with accepted safety practices. Samples should be diluted at least 1:400 with Sample Diluent A and can be further serially diluted if relative quantification is needed by the user. Positive and Negative Controls do NOT need to be diluted after reconstitution. Refer to the datasheet which shows typical results observed for the Positive Control, Negative Control, serum from uninfected individuals, and serum from SARS-CoV-2 patients. When using the cutoff criteria described below to determine if a sample is positive for anti-CoV-2 Spike Protein antibodies, samples diluted 1:400 must be compared to the undiluted Negative Control.
NOTE: Sample storage/handling, including heat-inactivation of samples, can potentially affect observed signals. Therefore, it is strongly recommended that in addition to the Positive and Negative Controls included with the kit, the user includes their own negative and positive patient samples as controls when running the assay in order to establish an appropriate cutoff value.
Add 100 µL of STOP Solution to each well. Shake gently for a few seconds.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
Limitations: Experimental cutoffs were determined by assaying a set of SARS-CoV-2 positive samples and uninfected donor serum collected prior to the SARS-CoV-2 pandemic.
NOTE: Positive samples were from donors with positive SARS-CoV-2 diagnosis, but IgM status for some samples may be negative due to timing of sample collection relative to disease onset.
Researchers can establish or modify this cutoff using additional samples. Positive or negative results from this assay should not be the sole basis for determining the infection status of a sample. A negative result can occur in SARS-CoV-2 patient samples due to:
posted August 2020
Protocol Id: 2224
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).
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.