SARS-CoV-2 (COVID-19) Spike RBD Antibody

• Catalog Number: orb1239994
• Category: Antibodies
• Description: SARS-CoV-2 (COVID-19) Spike RBD Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC, WB
• Reactivity: Virus
• Isotype: IgG
• Immunogen: Anti-SARS-CoV-2 (COVID-19) Spike RBD antibody (orb1239994) was raised against a peptide corresponding to 19 amino acids near the carboxy terminus of SARS-CoV-2 (COVID-19) Spike glycoprotein RBD. The immunogen is located within the last 50 amino acids of SARS-CoV-2 (COVID-19) Spike protein RBD.
• Concentration: 1 mg/mL
• Dilution range: WB: 2-5 μg/mL; IF: 20 μg/mL; IHC: 0.5 μg/mL.Antibody validated: Western Blot in human samples; Immunohistochemistry and Immunofluorescence in human samples. SARS-CoV-2 (COVID-19) Spike RBD Antibody can be used for the detection of SARS-CoV-2 (COVID-19) Spike protein in ELISA and WB. It will detect 4 ng of free peptide at 1 μg/mL. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• Target: S
• UniProt ID: P0DTC2
• NCBI: QHD43416
• Storage: SARS-CoV-2°COVID-19) Spike RBD antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: SARS-CoV-2 (COVID-19) Spike RBD antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: SARS-CoV-2 (COVID-19) Spike RBD Antibody: Severe a
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

ELISA Validation with SARS-CoV-2 (COVID-19) Spike RBD Recombinant Protein. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD antibody, orb1239994 (1 µg/mL). A direct ELISA was performed using SARS-CoV-2 (COVID-19) Spike RBD recombinant protein (orb1231588) as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike RBD antibody as the capture.

Detection of SARS-CoV-2 Variant Proteins with Spike RBD Antibodies by Direct ELISA. Coating Antigen: SARS-CoV-2 full length spike proteins, including WT, UK variant (B.1.1.7), SA variant (B.1.135) and Brazil (P.1). Dilution: 10-3000 ng/mL. Incubate at 4 ˚C overnight. Detection Antibodies: SARS-CoV-2 Spike RBD Antibody, orb1239994, 2 µg/mL, incubate at RT for 1 hr. Secondary Antibodies: Goat anti-rabbit HRP at 1:20000, incubate at RT for 1 hr. Immunogen region of antibody (orb1239994) includes site 501N that was mutated in all three variants. orb1239994 has low binding affinity for all three variants as compared to WT.

Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike RBD in COVID-19 Patient Lung. Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti-SARS-CoV-2 (COVID-19) Spike RBD antibody (orb1239994, 0.5 µg/mL). Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 ˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Strong signal of SARS-COV-2 Spike RBD protein was observed in macrophage of COVID-19 patient lung, but not in non-COVID-19 patient lung.

Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD Recombinant Protein. Loading: 30 ng per lane of SARS-CoV-2 (COVID-19) Spike RBD recombinant protein, orb1231588. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, orb1239994, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 µg/mL and Lane 2: 2 µg/mL.

Immunofluorescence Validation of SARS-CoV-2 (COVID-19) Spike RBD in 293 Transfected Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed Spike transfected 293 cells labeling SARS-CoV-2 (COVID-19) Spike RBD with orb1239994 at 20 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Overexpression Validation in Spike Transfected 293 Cells. Loading: 10 µg per lane of 293 cell lysate. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, orb1239994 (4 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: WT 293 cells and Lane 2: SARS-CoV-2 Spike overexpressed 293 cells.

Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD+SD1 Recombinant Protein. Loading: 30 ng per lane of SARS-CoV-2 (COVID-19) Spike RBD+SD1 recombinant protein, orb1231587. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, orb1239994, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 µg/mL and Lane 2: 2 µg/mL.

ELISA Validation with SARS-CoV-2 (COVID-19) Spike RBD+SD1 Recombinant Protein. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD antibody, orb1239994 (1 µg/mL). A direct ELISA was performed using SARS-CoV-2 (COVID-19) Spike RBD+SD1 recombinant protein (orb1231587) as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike RBD antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 8 ng/mL to 1000 ng/mL.

Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD+SD1+SD2 Recombinant Protein. Loading: 30 ng per lane of SARS-CoV-2 (COVID-19) Spike RBD+SD1+SD2 recombinant protein, orb1231586. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, orb1239994, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 µg/mL and Lane 2: 2 µg/mL.

ELISA Validation with SARS-CoV-2 (COVID-19) Spike Recombinant Protein. Antibodies: SARS-CoV-2 (COVID-19) Spike antibody, orb1239994 (1 µg/mL). A direct ELISA was performed using SARS-CoV-2 (COVID-19) Spike S1 recombinant protein (orb1227612) as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike RBD antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 16 ng/mL to 2000ng/mL.

Western Blot Validation with SARS-CoV-2 (COVID-19) Spike Recombinant Protein. Loading: 50 ng per lane of SARS-CoV-2 (COVID-19) Spike S1 recombinant protein (orb1227612). Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, orb1239994, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 2.5 µg/mL and Lane 2: 5 µg/mL.