Revision 4

#37322Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

H

UniProt ID:

#P0DTC2

Entrez-Gene Id:

#43740568

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
Spike Protein Coated Microwells II 87455 96 tests +4C
Anti-Human IgM, HRP-linked Antibody (ELISA Formulated) 15794 1 ea Red (Lyophilized) +4C
Sample Diluent A 71637 25 ml +4C
HRP Diluent 13515 11 ml Red +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 #37322 Positive Control 60497 1 ea +4C
ELISA Kit #37322 Negative Control 33807 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 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).

  1. Zhou, P. et al. (2020) Nature 579, 270-3.
  2. Tortorici, M.A. and Veesler, D. (2019) Adv Virus Res 105, 93-116.
  3. Li, F. et al. (2006) J Virol 80, 6794-800.
  4. Li, F. (2016) Annu Rev Virol 3, 237-61.
  5. Shang, J. et al. (2020) Nature 581, 221-4.
  6. Wrapp, D. et al. (2020) Science 367, 1260-3.
  7. Yan, R. et al. (2020) Science 367, 1444-8.
  8. Yuan, Y. et al. (2017) Nat Commun 8, 15092.
  9. 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

    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.

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    Revision 4
    #37322

    SARS-CoV-2 Spike Protein Serological IgM ELISA Kit

    SARS-CoV-2 Spike Protein Serological IgM ELISA Kit: Image 1 Expand Image
    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.

    SARS-CoV-2 Spike Protein Serological IgM ELISA Kit: Image 2 Expand Image
    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.
    SARS-CoV-2 Spike Protein Serological IgM ELISA Kit: Image 3 Expand Image
    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.
    SARS-CoV-2 Spike Protein Serological IgM ELISA Kit: Image 4 Expand Image
    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.