• Product NameAnti-Nucleocapsid (N) Rabbit Monoclonal Antibody
• Brief DescriptionAntibody
• Clone No.V400-G1
• ApplicationsWB, ELISA
• Species ReactivitySARS-CoV-2
• Immunogen DescriptionRecombinant SARS-CoV-2 nucleocapsid protein
• Alternative Names    Severe acute respiratory syndrome coronavirus 2 nucleocapsid; 2019 novel coronavirus nucleoprotein, SARS-CoV-2 NP, SARS-CoV-2 N protein
• Accession No. UniProt: P59595
• UniprotP59595
• Calculated MW 46 kDa
• Formulation Supplied in PBS (pH 7.5) containing 50 ?g/mL BSA, 50% glycerol, and 0.01% sodium azide
• Storage Store at -20˚C

• Western blotting (WB): 1:1,000; ELISA: 1:10,000

The single-stranded, plus-sense severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome encodes nonstructural replicase polyproteins as well as structural proteins such as spike (S), nucleocapsid (N), membrane (M) and envelop (E) proteins (Zhou et al., 2020). N protein is abundantly expressed and highly immunogenic during SARS-COV-2 infection (Cong et al., 2020). The middle or C-terminal region of the N protein has been shown to elicit antibody production during the immune response (Dutta et al., 2020). Although the surface electrostatic potential characteristics of SARS-CoV-2 N is different from other coronavirus N proteins (Kang et al., 2020), the sequences are conserved among these proteins (Dutta et al., 2020). In addition to S protein, N protein is also considered as a leading target antigen for vaccine development (Chen et al., 2020; Dutta et al., 2020).

• Figure 1. Antibody reactivity and specificity test. Three positive and two negative human nasopharyngeal swab specimens (NP Swabs), and gamma-irradiated SARS-CoV-2 (nCoV), SARS-CoV (SARS), and MERS were lysed in Viral Lysis Buffer (Cat# VL101) and blotted with rabbit monoclonal anti-N primary antibody (Cat# VYN4, 1:2,000 dilution). HRP-conjugated highly cross-adsorbed goat anti-rabbit IgG secondary antibody (Cat# 202) was used as the secondary antibody. The blot was then incubated using PiQTM ECL Substrate Kit (Cat# 639) and imaged using a chemiluminescence digital imager. Note that 1) NP Swab POS No.3 has very low viral load; 2) There were no cross-reactivity of the antibody with either SARS or MERS.
• Figure 3. Chemiluminescent ELISA using recombinant nucleocapsid protein (rN). Different concentrations of rN (Cat# VPN1) lysed in Viral Lysis Buffer (Cat# VL101) were coated on microplate wells. Rabbit monoclonal anti-N primary antibody (Cat# VYN4, 1:10,000 dilution) and HRP-conjugated highly cross-adsorbed goat anti-rabbit IgG secondary antibody (Cat# 202, 1:20,000) were used to detect SARS-CoV-2 N protein. ShQTM ECL reagents (Cat# 707) were used to generate chemiluminescent signals. RLU, relative light unit. ΔRLU = RLU of the well with primary antibody - RLU of the corresponding well without primary antibody, all else being equal.
• Figure 2. Chemiluminescent ELISA using SARS-CoV-2. Gamma-irradiated SARS-CoV-2 viruses were lysed in Viral Lysis Buffer (Cat# VL101) and coated (150 μL) on microplate wells. Rabbit monoclonal anti-N primary antibody (Cat# VYN4, 1:10,000 dilution) and HRP-conjugated highly cross-adsorbed goat anti-rabbit IgG secondary antibody (Cat# 202, 1:20,000) were used to detect SARS-CoV-2 N protein. PiQTM ECL (Cat# 636) reagents were used to generate chemiluminescent signals. RLU, relative light unit. ΔRLU = RLU of the well with primary antibody - RLU of the corresponding well without primary antibody, all else being equal.
• Figure 4. Chemiluminescent ELISA using contrived human nasopharyngeal (NP) swab specimens. Different concentrations of recombinant nucleocapsid protein (rN) (Cat# VPN1) were spiked in RT-PCR-confirmed negative human NP swabs and lysed in Viral Lysis Buffer (Cat# VL101). The lysates were coated on microplate wells, and rabbit monoclonal anti-N primary antibody (Cat# VYN4, 1:10,000 dilution) and HRP-conjugated highly cross-adsorbed goat anti-rabbit IgG secondary antibody (Cat# 202, 1:20,000) were used to detect SARS-CoV-2 N protein. ShQTM ECL reagents (Cat# 707) were used to generate chemiluminescent signals. RLU, relative light unit. ΔRLU = RLU of the well with primary antibody - RLU of the corresponding well without primary antibody, all else being equal.

• Flow Cytometry (FCM) General Protocol
• Immunofluorescence(IC)
• Immunohistochemistry(Polymer)
• Western Blotting