Revision 2

#76349Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

Vir

UniProt ID:

#P0DTC2

Entrez-Gene Id:

#43740568

Cell Signaling Technology

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color
FastScan ELISA Microwell Strip Plate, 96 Well 53257 96 tests
SARS-CoV-2 Spike Protein Mouse Capture mAb 37174 1 ea Green (Lyophilized)
SARS-CoV-2 Spike Protein Rabbit HRP-linked mAb 72574 1 ea Red (Lyophilized)
FastScan ELISA Capture Antibody Diluent 16076 3 ml Green
FastScan ELISA HRP Antibody Diluent 28120 3 ml
TMB Substrate 7004 11 ml
STOP Solution 7002 11 ml
Sealing Tape 54503 1 ea
ELISA Wash Buffer (20X) 9801 25 ml
FastScan ELISA Cell Extraction Buffer (5X) 69905 10 ml
FastScan ELISA Cell Extraction Enhancer Solution (50X) 25243 1 ml
FastScan ELISA Kit #76349 Positive Control Type 2 41492 1 ea

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The FastScan SARS-CoV-2 Spike Protein ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of SARS-CoV-2 spike protein. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with SARS-CoV-2 spike protein in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of SARS-CoV-2 spike protein.

*Antibodies in kit are custom formulations specific to kit.

IMPORTANT: This FastScan ELISA Kit requires 4 washes at Step 6 of the protocol.

Specificity/Sensitivity

The FastScan SARS-CoV-2 Spike Protein ELISA Kit detects endogenous levels of SARS-CoV-2 spike protein, but does not cross-react with spike proteins from SARS or MERS coronaviruses, as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. This ELISA kit will detect full-length SARS-CoV-2 spike protein and SARS-CoV-2 spike protein fragments containing both the RBD and the S1-NTD regions, as shown in Figure 2 (one ELISA antibody recognizes a region in the S1-NTD of SARS-CoV-2 spike protein, while the other antibody targets a region in the RBD).

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.
    FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.
    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.

    Revision 2
    #76349

    FastScan SARS-CoV-2 Spike Protein ELISA Kit

    FastScan™ SARS-CoV-2 Spike Protein ELISA Kit: Image 1 Expand Image
    Figure 1. The FastScan SARS-CoV-2 Spike Protein ELISA Kit #76349 detects SARS-CoV-2 but not SARS-CoV or MERS-CoV spike proteins. The absorbance at 450 nm using the FastScan SARS-CoV-2 Spike Protein ELISA Kit #76349 for cell extracts (0.05 mg/mL) from 293T cells that were mock transfected or transiently transfected with expression constructs encoding Myc/DDK-tagged SARS-CoV-2 spike, Myc/DDK-tagged SARS-CoV spike, or Myc/DDK-tagged MERS-CoV spike, is shown in the left panel. The corresponding western blots of these cell extracts are shown in the right panel using SARS-CoV-2 Spike Protein (S1-NTD) (E7M5X) Mouse mAb #42172 (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower) (lane 1: mock transfected, lane 2: Myc/DDK-tagged SARS-CoV-2 spike, lane 3: Myc/DDK-tagged SARS-CoV spike, lane 4: Myc/DDK-tagged MERS-CoV spike).
    FastScan™ SARS-CoV-2 Spike Protein ELISA Kit: Image 2 Expand Image
    Figure 2. The relationship between the concentration of SARS-CoV-2 Spike (trimeric) (16-1208) Recombinant Protein (8xHis-Tag) #65444, SARS-CoV-2 Spike RBD (318-541) Recombinant Protein (8xHis-Tag) #48801, SARS-CoV-2 Spike S1-NTD (16-316) Recombinant Protein (8xHis-Tag) #88587 and the absorbance at 450 nm using the FastScan SARS-CoV-2 Spike Protein ELISA Kit #76349 is shown in the figure. As indicated in the Specificity/Sensitivity section, one of the ELISA antibodies recognizes a region in the S1-NTD of SARS-CoV-2 spike protein, while the other antibody targets a region in the RBD; therefore, this ELISA kit will only recognize full-length SARS-CoV-2 spike protein and SARS-CoV-2 spike protein fragments containing both the RBD and the S1-NTD regions.