21MAk-S2N-B12N

Mouse monoclonal antibody (cl. B12N/7F10) to SARS-CoV-2 nucleocapsid protein (N)

Description: Mouse monoclonal antibody (clone B12N) to SARS-CoV-2.

Clone: B12N

Original clone name: 7F10

Host: mouse

Subclass: IgG1

Immunogen: SARS-CoV-2 full-length nucleocapsid protein from S. cerevisiae (Baltymas product code: 20-S2N-ScA-F).

Tested applications: ELISA, Western blot (WB), imunofluorescence (IF).

Image: SDS-PAGE and Western blotting using monoclonal antibody B12N/7F10 to SARS-CoV-2 nucleocapsid protein (cat. code 21MAk-S2N-B12N), 0.1 µg/ml, followed by secondary goat anti-mouse IgG antibody conjugated to HRP.

M – PageRuler Prestained Protein Ladder, 5 µl/well

1 – SCoV2-rN, SARS-CoV-2 nucleoprotein from S. cerevisiae (Baltymas product 20-S2N-ScB-F), 1 µg/well

2 – SCoV2-DN-rN, SARS-CoV-2 nucleoprotein C-terminal domain from S. cerevisiae (Baltymas product 21-S2delNN-ScB-F), 1 µg/well

3 – S. cerevisiae lysate (negative control)

4 – SCoV2-rS, SARS-CoV-2 Spike glycoprotein full-length trimeric ectodomain from mammalian HEK293 cells (negative control, Baltymas product 20-S2S-THs-F), 1 µg/well

5 – SCoV2-RBD,  SARS-CoV-2 Spike glycoprotein Receptor Binding Domain from mammalian HEK293 cells (negative control, Baltymas product 20-S2RBD-RHs-F), 1 µg/well.

Proteins were separated in 12% SDS-PAGE.

Chromogenic detection was performed by 4-Chloro-1-naphthol.

Image 2: Western blot using monoclonal antibody B12N/7F10 to SARS-CoV-2 nucleocapsid protein (Cat. code 21MAk-S2N-B12N), 5 µg/ml, followed by secondary anti-mouse antibody conjugated to HRP.

M – PageRuler™ Prestained Protein Ladder, 10 to 180 kDa, Thermo Fisher Scientific, 1 – SARS-CoV-2 nucleocapsid protein (20-S2N-ScA-F), 1 µg, 2 – SARS-CoV-1 nucleocapsid protein (20-S1N-ScB-F), 1 µg.   Proteins were separated in 12% SDS-PAGE. Chromogenic detection was performed by TMB for membranes.

Note: The antibody B12N/7F10 reacts with both SARS-CoV and SARS-CoV-2 nucleoproteins.

Graph: ELISA titration curves.

ELISA titration curves using monoclonal antibody B12N to SARS-CoV-2 nucleocapsid protein (Cat. code 21MAk-S2N-B12N), followed by secondary anti-mouse antibody conjugated to HRP.   1 – SARS-CoV-2 nucleocapsid protein (20-S2N-ScB-F), 2 – SARS-CoV-1 nucleocapsid protein (20-S1N-ScB-F). 3 – SARS-CoV-2 nucleocapsid (Omicron BA.5 variant) protein (22-S2NOM5-ScB-F)

Apparent dissociation constants of B12N:

SARS-CoV-2 nucleocapsid protein (20-S2N-ScB-F) – 3,04 × 10^-10 ± 9,11 × 10^-12 M

SARS-CoV-1 nucleocapsid protein (20-S1N-ScB-F) – 1,22 × 10^-10 ± 1,87 × 10^-11 M

SARS-CoV-2 nucleocapsid (omicron BA.5 variant) protein (22-S2NOM5-ScB-F) – 8,1 × 10^-11 ± 4,32 × 10^-12 M

Recommended starting dilutions: WB 1/200-1/1000; ELISA 1/1000-1/10000. Optimal concentrations should be determined by the end user.

Immunofluorescence photos: Based on an indirect immunofluorescence test, monoclonal antibody B12N/7F10 (10 µg/ml) recognizes ancestor strain SARS-CoV-2 virus in Vero E6 cells.

Uninfected cells (negative control)	Infected cells

Purity: Protein A purified

Form: Sterile-filtered PBS, pH 7.2

Concentration: 1 mg/ml

Storage conditions: This product is stable at +2 – +8°C for 2-3 weeks. For a long-term storage the antibody can be stored at -20°C or -80°C (avoid freeze / thaw cycle). Alternatively, in 50% glycerol at -20°C.

Conditions of shipping: Blue ice (0-8°C). Shipment cost to Europe: 100 €.

Additional info: The epitope for SARS-CoV-2 nucleocapsid protein recognised by monoclonal antibody B12N is located at the N-terminus, between 1 and 94 aa (Protein Accession No. in Uniprot:  P0DTC9 (NCAP_SARS2)).

Availability: can be prepared for shipment in 1 week.

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