SARS-CoV-2 (COVID-19) Spike S2 Antibody [5E6]

• Catalog Number: orb1238677
• Category: Antibodies
• Description: SARS-CoV-2 (COVID-19) Spike S2 Antibody [5E6]
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: [5E6]
• Tested applications: ELISA, IF, IHC-P, WB
• Reactivity: Virus
• Isotype: IgG1
• Immunogen: Anti-SARS-CoV-2 (COVID-19) Spike S2 antibody (orb1238677) was raised against a peptide corresponding to 20 amino acids near the carboxy terminus of SARS-CoV-2 (COVID-19) Spike glycoprotein. The immunogen is located within the last 50 amino acids of SARS-CoV-2 (COVID-19) Spike protein.
• Concentration: 1 mg/mL
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• Target: S
• UniProt ID: P0DTC2
• NCBI: QHD43416
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: SARS-CoV-2 (COVID-19) Spike Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: SARS-CoV-2 (COVID-19) Spike S2 Antibody: Severe ac
• Note: For research use only
• Application notes: WB: 0.2 μg/mL; IF: 20 μg/mL. IHC: 0.5 μg/mL; Antibody validated: Immunofluorescence and Western blot in human samples. Immunohistochemistry in COVID-19 patient samples. SARS-CoV-2 (COVID-19) Spike S2 antibody can be used for the detection of SARS-CoV-2 (COVID-19) Spike full length protein in ELISA. It will detect 4 ng of free peptide at 1 μg/mL. orb1238677 cannot be used for detection of SARS-CoV-2/SARS-CoV Spike S1, Spike ECD, Spike S2 ECD proteins. All other applications and species not yet tested.
• Expiration Date: 12 months from date of receipt.

Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike in COVID-19 Patient Lung.Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti-SARS-CoV-2 (COVID-19) Spike S2 antibody (orb1238677, 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-mouse IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Strong spike protein signal was observed in macrophages of COVID-19 patient lung, but not in non-COVID-19 patient lung.

Overexpression Validation in Spike Transfected 293 Cells Loading: 5 μg per lane of 293 cell lysate. Antibodies: SARS-CoV-2 (COVID-19) Spike S2, orb1238677, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10, 000 dilution. Lane 1: 0.1 μg/mL and.Lane 2: 0.2 μg/mL

Immunofluorescence Validation of SARS-CoV-2 (COVID-19) Spike in 293T Cells.Immunofluorescent analysis of 4% paraformaldehyde-fixed 293T cells labeling SARS-CoV-2 (COVID-19) Spike S2 with orb1238677 at 20 μg/mL, followed by goat anti-mouse IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

ELISA Test. Antibodies: SARS-CoV-2 (COVID-19) Spike S2 antibody, orb1238677 (1 μg/mL). A direct ELISA was performed using immunogen or control peptide as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike antibody as the capture antibody. Secondary: Goat anti-mouse IgG HRP conjugate at 1:5.000 dilution. Detection range is from 0.3 ng/mL to 1000 ng/mL.