Revision 6

#37322

Store at +4C

SARS-CoV-2 Spike Protein Serological IgM ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H

UniProt ID:
#P0DTC2

Entrez-Gene Id:
#43740568

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Product Includes Product # Quantity Color Storage Temp
Spike Protein Coated Microwells II8745596 tests+4C
Anti-Human IgM, HRP-linked Antibody (ELISA Formulated)157941 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 #37322 Positive Control604971 ea+4C
ELISA Kit #37322 Negative Control338071 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.

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 Protein Serological IgM ELISA Kit is a solid-phase ELISA that detects binding of human IgM to full-length ectodomain SARS-CoV-2 spike trimeric protein (S-protein). Trimeric spike protein has been coated onto microwells. After incubation with sample, human IgM specific for trimeric spike protein is captured on the plate. The wells are then washed to remove unbound material. Anti-Human IgM, HRP-linked antibody is then used to recognize the bound IgM. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of IgM specific for spike protein.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

The SARS-CoV-2 Spike Protein Serological IgM ELISA Kit detects endogenous levels of human IgM directed to full-length ectodomain SARS-CoV-2 spike trimeric protein (S-protein).

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).

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.

限制使用

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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

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.

Revision 6

Cell Signaling Technology Logo
Patient Testing: Patient samples were tested using the SARS-CoV-2 Spike Protein Serological IgM ELISA Kit #37322. Serum or plasma was obtained from donors who had been diagnosed with SARS-CoV-2 (diagnosed positive n=49*) or from presumed uninfected donors collected prior to the SARS-CoV-2 outbreak (presumed negative n=86). Samples were heat-inactivated (56°C for 30 min) and diluted 1:400 prior to running the assay, as described in the protocol. Samples were considered positive, negative, or inconclusive based on the criteria described in the “Data Analysis” section of the protocol for the kit. Positive percent agreement (PPA) and negative percent agreement (NPA) were calculated from these data.

*Positive reference samples were from patient donors with a positive SARS-CoV-2 diagnosis. However, a positive SARS-CoV-2 diagnosis will not always correlate with a positive response in the IgM Serological ELISA as differences in disease severity and timing of sample collection relative to disease onset will affect presence and abundance of IgG, IgA, and IgM in the reference sample.

Note: We are continuing to test more samples as available. For the most up-to-date set of data, always refer to the product page for #37322 on the website.

ELISA Image 1: SARS-CoV-2 Spike Protein Serological IgM ELISA Kit
Intra-Assay Precision: Three different serum samples were each tested in 16 replicates using a single assay kit of SARS-CoV-2 Spike Protein Serological IgM ELISA #37322. Intra-assay CV (%) was calculated for each sample, and each replicate was correctly identified as being positive or negative when compared to the Negative Control using the cutoff criteria described in the attached protocol.
ELISA Image 2: SARS-CoV-2 Spike Protein Serological IgM ELISA Kit
Inter-Assay Precision: Six different assay kits from one lot of material were tested using 3 different serum samples run in duplicate wells and Positive and Negative Controls run in 4 replicate wells. Inter-assay CV (%) was calculated for each sample, and each assay kit correctly identified the samples as being positive or negative when compared to the Negative Control using the cutoff criteria described in the attached protocol.
ELISA Image 3: SARS-CoV-2 Spike Protein Serological IgM ELISA Kit
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.

Revision 6

Cell Signaling Technology Logo
Patient Testing: Patient samples were tested using the SARS-CoV-2 Spike Protein Serological IgM ELISA Kit #37322. Serum/plasma samples were heat-inactivated (56°C for 30 min) and diluted 1:400 prior to running the assay, as described in the protocol. Background subtracted absorbance values at 450 nm are plotted for each serum/plasma sample, with values corresponding to samples obtained from donors who had been diagnosed with SARS-CoV-2 (diagnosed positive n=49*) on the right, and values corresponding to samples from presumed uninfected donors collected prior to the SARS-CoV-2 outbreak (presumed negative n=86) on the left. Also shown is the cutoff line above which samples are considered positive (blue line), and the negative cutoff line (red line), below which samples are considered negative. Cutoff values were calculated as described in the “Data Analysis” section of the protocol for the kit.

*Positive reference samples were from patient donors with a positive SARS-CoV-2 diagnosis. However, a positive SARS-CoV-2 diagnosis will not always correlate with a positive response in the IgM Serological ELISA as differences in disease severity and timing of sample collection relative to disease onset will affect presence and abundance of IgG, IgA, and IgM in the reference sample.

ELISA Image 4: SARS-CoV-2 Spike Protein Serological IgM ELISA Kit
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.