Anti SARS-CoV Nucleocapsid (N) Protein

Immunofluorescence of Rabbit Anti-SARS-CoV Nucleocapsid (N) Antibody.
Tissue: human Liver ductal organoids.
Fixation: 4% PFA.
Permeabilization: 0.25% Triton X-100.
Antigen retrieval: not required.
Primary antibody: Rabbit Anti-SARS-CoV (N) Antibody and Mouse Anti-E-Cadhedrin Antibody at 1:500 overnight at 2-8C.
Secondary antibody: Donkey Anti-Rabbit IgG CY3 Conjugated; Donkey Anti-Mouse IgG AlexaFluor 488 Conjugated for 1hr at RT.
Nuclear Counterstain: DAPI.
Staining showing Mock and Infected tissue: SARS-CoV Red signal, E-Cadhedrin green signal, with DAPI (blue) nuclear counterstain.
[Zhao et al. (2020)]

Immunofluorescence of Rabbit Anti-SARS-CoV Nucleocapsid (N) Antibody.
Tissue: human Liver ductal organoids.
Fixation: 4% PFA.
Permeabilization: 0.25% Triton X-100.
Antigen retrieval: not required.
Primary antibody: Rabbit Anti-SARS-CoV (N) Antibody and Mouse Anti-E-Cadhedrin Antibody at 1:500 overnight at 2-8C.
Secondary antibody: Donkey Anti-Rabbit IgG CY3 Conjugated; Donkey Anti-Mouse IgG AlexaFluor 488 Conjugated for 1hr at RT.
Nuclear Counterstain: DAPI.
Staining of Infected and Merged: SARS-CoV Red signal, E-Cadhedrin green signal, with DAPI (blue) nuclear counterstain.
[Zhao et al. (2020)]

Western Blot of Rabbit Anti-SARS CoV Nucleocapsid (N) Protein Antibody.
Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500).
Lane 2: SARS CoV Nucleocapsid (N) Protein [20ng].
Primary Antibody: Anti-SARS CoV Nucleocapsid (N) Protein Antibody at 1.0μg/mL overnight at 2-8C.
Secondary Antibody: Goat Anti-Rabbit IgG HRP (p/n 611-1302) at 1:40000 for 30mins at RT.
Block: BlockOut Buffer (p/n MB-073).
Predicted MW: ~46kDa.
Observed MW: ~48kDa.

Western blot using Rockland's Protein A Purified anti-SARS CoV Nucleocapsid (N) protein antibody shows detection of a 46-kDa band corresponding to the protein. Approx. 100 ng of protein was loaded for SDS-PAGE and transferred onto nitrocellulose. The blot was incubated with a 1:5,000 dilution of the antibody at room temperature for 1 h followed by detection using IRDye^TM800 labeled Goat-a-Rabbit IgG [H&L] (611-132-122) diluted 1:10,000. The fluorescence image was captured using the Odyssey^(R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.

Western Blot of Rabbit Anti-SARS-CoV (N) Antibody tested on Caco-2 Cell Lysates before (mock), 24, and 48 hours post infection (hpi).
Primary Antibody: Anti-SARS-CoV Nucleocapsid (N) Antibody.
Observed MW: ~46kDa in infected cells.
[Courtesy of Sebastian Weigang - Kochs Labs]

Immunofluorescence assay using Rabbit Anti-SARS-CoV Nucleocapsid (N) Antibody, showing viral protein synthesis. VeroE6 cells were infected with rSARS-CoV-2. Mock: non-infected cells.
At 48 h.p.i, cells were fixed and prepared for immunofluorescence staining with primary antibodies directed against double-stranded RNA (dsRNA) [green] and SARS-CoV Nucleocapsid (N) [red]. Nuclear Counterstain DAPI [blue].

Immunohistochemistry results showing positive staining in placenta tissue using Anti-SARS-CoV Nucleocapsid (N) Antibody.

Immunohistochemistry of Rabbit Anti-SARS CoV Nucleocapsid Antibody.
Pre-treated tissue: (A) human lung control, (B) interferon stimulated genes (ISG) low, (C) ISG high human COVID-19 lung: interferon stimulated genes (ISGs). [One pattern showes high expression of interferon stimulated genes (ISGs) and cytokines, high viral loads and limited pulmonary damage, the other pattern showes severely damaged lungs, low ISGs, low viral loads and abundant immune infiltrates.]
Primary Antibody: Anti-SARS-CoV at 1:6400.
Staining and Detection with Bond Polymer Refine Red Detection.
[Courtesy of Mertz K. Profiles of COVID-19 lungs.]

ELISA results of Rabbit Anti-SARS-CoV Nucleocapsid (N) Protein. Each well was coated in duplicate with 0.5μg/mL of SARS CoV-1 [Red Line], SARS CoV-2 Nc [Green Line], MERS CoV N [Blue Line]. The starting dilution of antibody was 2μg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. SARS CoV-1N IC50: 11ng/mL, SARS CoV-2N IC50: 15ng/mL. Assay performed using Goat Anti-Rabbit IgG HRP conjugated (p/n 611-1302) and TMB substrate (p/n TMBE-1000).

Immunofluorescence of Rabbit Anti-SARS CoV (M) Protein Antibody.
Cells: Caco-2 cells 24 hours post-infection.
Fixation: 4% PFA.
Permeabilization: 0.3% Triton X-100.
Blocking: 5% fetal calf serum/PBS.
Primary Antibody: Anti-SARS-CoV nucleocapsid (N) protein at 1:1000 for 1 hour at RT.
Staining: N-Protein [Red], F-actin [White], DAPI, [Blue].
Imaged: Zeiss LSM800 microscope.
[Images Courtesy of AG Robert Grosse, Institute of Experimental and Clinical Pharmacology I, University of Freiburg/Svenja Ulferts and AG Georg Kochs Labs/Sebastian Weigang]. [Bouhaddou M et al. 2020]