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23733
SignalScan™ Peptide Mix (SARS-CoV-2)
Proteomic Analysis Products
AQUA Peptide

SignalScan™ Peptide Mix (SARS-CoV-2) #23733

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SignalScan™ Peptide Mix (SARS-CoV-2): Image 1

Product ion chromatograms, visualized in Skyline, of peptides from the human ACE2 protein (top) and the SARS-CoV-2 Nucleocapsid protein (bottom), derived from virally-infected A549 cells. Using a standard data-dependent analysis (DDA) LCMS/MS method (left panels) results in no signal or poor peak shape due to instrument duty cycle. A Stable Isotope Standard Assisted Scanning (SISAS) method (right panels) results in consistently detected peptides with quality peak shapes resulting in confident peptide and protein quantitaton.

SignalScan™ Peptide Mix (SARS-CoV-2): Image 2

SignalScan™ LCMS Analysis. SignalScan™ uses a stable isotope standard assisted scanning (SISAS) principle to quantitate targeted peptides without retention time scheduling and with greater sensitivity than untargeted DDA approaches. The heavy standard and the matching endogenous peptide co-elute in the chromatogram but are resolved in the MS1 scan by a mass difference of +8 or +10Da.  As peptides enter the mass spectrometer, only MS1 precursor scans are performed until an m/z matching one of the heavy SIS peptides is found. Then a fast, low-resolution MS2 scan event fragments the SIS precursor ion and matches its most abundant fragments to a list of known values. This confirms the identity of the SIS peptide and triggers analysis of the matching endogenous peptide. The light peptide is fragmented and a series of MS2 scans are collected with parameters optimized for high-resolution quantitation. Mutliple fragment intensities are monitored as the peptide elutes, just as in a parallel-reaction-monitoring experiment. The areas under the peak curves are integrated in Skyline and the relative area changes can be calculated for each treatment condition.

SignalScan™ Peptide Mix (SARS-CoV-2): Image 3

SignalScan™ Workflow Steps. The SignalScan™ Peptide Mix (SARS-CoV-2) is compatible with any human sourced biological material. Extract proteins from the tissue and digest them into peptides using any standard trypsin-based protocol. Spike in 1 pmol of the heavy Stable Isotope Standard (SIS) peptides provided prior to LC-MS/MS analysis.

Product Description

The SignalScan™ Peptide Mix (SARS-CoV-2) enables targeted analysis of selected peptides in human-sourced cells or tissues. Peptides and phosphopeptides derived from SARS-CoV-2 spike and nucleocapsid proteins as well as nine human host response proteins are included. This mixture contains 96 pmol each of 25 heavy-labeled (13C, 15N) Stable Isotope Standard (SIS) peptides, quantified by amino acid analysis (AAA). Inclusion of SIS peptides enhances the sensitivity of the LCMS assay by acting as a trigger for targeted MS/MS analysis. Targeted assays ensure that peptides present in the biological sample of interest are consistently observed and sensitivity is maximized. SIS-assisted scanning provides the sensitivity of a parallel reaction monitoring (PRM) assay without use of rigid retention time windows. Full peptide list, detailed protocol, and files for method set-up and data analysis are available for download at http://media.cellsignal.com/www/zip/proteomics/signalscan-peptide-mix.zip.

Storage

This product is stable for 12 months when stored at -20ºC. Aliquot to avoid multiple freeze/thaw cycles.

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). The cell’s subsequent host response to SARS-CoV-2 infection involves complex signaling cascades, observable through perturbations in protein phosphorylation patterns (8,9) that lead to interferon and cytokine production (10).

  1. Zhou, P. et al. (2020) Nature 579, 270-273.
  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, 237-261.
  5. Shang, J. et al. (2020) Nature 581, 221-224.
  6. Wrapp, D. et al. (2020) Science 367, 1260-1263.
  7. Yan, R. et al. (2020) Science 367, 1444-1448.
  8. Davidson, A.D. et al. (2020) Genome Med 12, 68.
  9. Bouhaddou, M. et al. (2020) Cell 182, 685-712.e19.
  10. Blanco-Melo, D. et al. (2020) Cell 181, 1036-1045.e9.

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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalScan is a trademark of Cell Signaling Technology, Inc.
SureQuant is a registered trademark of Thermo Fisher Scientific Inc.
To Purchase # 23733S
Product # Size Price
23733S
96 µl  (96 assays) $ 499