Western blot analysis of extracts from mock infected Vero-E6 cells (lane 1), SARS-CoV-2-infected Vero-E6 cells (lane 2), and isolated SARS-CoV-2 virions (lane 3), using SARS-CoV-2 Nucleocapsid Protein Antibody (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). SARS-CoV-2 virions and infected Vero-E6 cells courtesy of Dr. Mohsan Saeed, National Emerging Infectious Diseases Laboratories, Boston University.
Western blot analysis of extracts from transgenic A549 cells stably expressing HA-tagged SARS-CoV-2 nucleocapsid (N) protein (A549-N protein) (lane 1) or HA-tagged SARS-CoV-2 envelope (E) protein (A549-E protein) (lane 2), using SARS-CoV-2 Nucleocapsid Protein Antibody (upper), HA-Tag (6E2) Mouse mAb #2367 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Absence of signal in A549-E protein cell extracts confirms specificity of the antibody for SARS-CoV-2 nucleocapsid (N) protein. Transgenic A549 cells were generously provided by Dr. Elena Piskounova, Weill Cornell Medicine.
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged SARS-CoV-2 nucleocapsid (N) protein (lane 2), using SARS-CoV-2 Nucleocapsid Protein Antibody (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
SARS-CoV-2 Nucleocapsid Protein Antibody recognizes endogenous levels of total SARS-CoV-2 nucleocapsid (N) protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly30 of SARS-CoV-2 nucleocapsid (N) protein. Antibodies are purified by protein A and peptide affinity chromatography.
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 relatively large and encodes up to 29 open reading frames (ORFs). These include ORF1a and ORF1b (further processed into 16 non-structural proteins), 9 accessory proteins, and 4 canonical structural proteins: spike (S), envelope (E), membrane (M), and nucleocapsid (N) (2). The nucleocapsid (N) protein is a phosphoprotein that plays a critical role in viral assembly (3,4). It binds to the viral genomic RNA, enabling its packaging into the ribonucleoprotein complex (RNP), and is the most abundant protein in the virion (5). Evidence from studies in SARS-CoV-2 and other coronaviruses suggest functions for nucleocapsid (N) protein beyond viral RNA packaging, including roles in viral replication (6) and suppressing host immune responses to viral infection (7).
Explore pathways + proteins related to this product.
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