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SARS-CoV-2 (2019-nCoV) Nucleoprotein/NP ELISA Kit

SB Sino Biological

Published: Apr 5, 2020 DOI: 10.21769/BioProtoc.5001 Views: 1243

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Background

Coronaviruses are enveloped viruses with a positive-sense RNA genome and with a nucleocapsid of helical symmetry. Coronavirus nucleoproteins localize to the cytoplasm and the nucleolus, a subnuclear structure, in both virus-infected primary cells and in cells transfected with plasmids that express N protein. Coronavirus N protein is required for coronavirus RNA synthesis, and has RNA chaperone activity that may be involved in template switch.Nucleocapsid protein is a most abundant protein of coronavirus. During virion assembly, N protein binds to viral RNA and leads to formation of the helical nucleocapsid. Nucleocapsid protein is a highly immunogenic phosphoprotein also implicated in viral genome replication and in modulating cell signaling pathways. Because of the conservation of N protein sequence and its strong immunogenicity, the N protein of coronavirus is chosen as a diagnostic tool.
Note: The use of this kit for natural samples need be validated by the end user due to the complexity of natural targets and unpredictable interference.

Materials and Reagents

  1. SARS-CoV-2 (2019-nCoV) Nucleoprotein/NP Microplate - 96 well polystyrene microplate (12 strips of 8 wells) coated with rabbit mAb antibody against
  2. SARS-CoV-2 (2019-nCoV) Nucleoprotein/NP
  3. SARS-CoV-2 (2019-nCoV) Nucleoprotein/NP Detection Antibody - 0.2 mg/ml of rabbit mAb antibody against SARS-CoV-2 (2019-nCoV) Nucleoprotein/NP conjugated to horseradish peroxidase (HRP) with preservatives
  4. SARS-CoV-2 (2019-nCoV) Nucleoprotein/NP Standard - Recombinant SARS-CoV-2 (2019-nCoV) Nucleoprotein/NP in a buffer with preservatives, lyophilized. The amount of standard is lot specific and indicated on the label of standard vial
  5. Wash Buffer Concentrate - 25 ml of a 20-fold concentrated solution of buffered surfactant with preservatives, stored for up to 1 week at 2-8 °C
  6. Dilution Buffer Concentrate - 8 ml of a 20-fold concentrated dilution buffer with preservatives, stored for up to 1 week at 2-8 °C
  7. 5× Sample Lysis Buffer - 6 ml, stored for up to 1 week at 2-8 °C
  8. Color Reagent A - 13 ml of stabilized hydrogen peroxide, stored for up to 1 month at 2-8 °C
  9. Color Reagent B - 13 ml of stabilized chromogen (tetramethylbenzidine), stored for up to 1 month at 2-8 °C
  10. Stop Solution - 8 ml of 2 N sulfuric acid, stored for up to 1 month at 2-8 °C
  11. Deionized or distilled water

Other supplies required
  1. Pipette tips
  2. Tubes for standard dilution
  3. Well plate cover or seals

Equipment

  1. Microplate reader capable of measuring absorbance at 450 nm
  2. Pipettes 
  3. Multi-channel pipette, squirt bottle, manifold dispenser, or automated microplate washer
  4. 500 ml graduated cylinder

Procedure

Copyright: © 2020 The Author(s); This is an open access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/).

Category

Bioinformatics and Computational Biology

Systems Biology > Genomics > Phylogenetics

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