ORF3a Antibody (2819D) [Unconjugated]

Western blot shows recombinant SARS-CoV-2 ORF3a peptide. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-SARS-CoV-2 ORF3a Monoclonal Antibody (Catalog # MAB107061) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for ORF3a at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

ORF3a was detected in immersion fixed HEK293 human embryonic kidney cell line transfected with SARS-CoV-2 (positive staining) and HEK293 human embryonic kidney cell line (non-transfected, negative staining) using Rabbit Anti-SARS-CoV-2 ORF3a Monoclonal Antibody (Catalog # MAB107061) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

ORF3a was detected in immersion fixed paraffin-embedded sections of human spleen infected with SARS-CoV-2 using Rabbit Anti-SARS-CoV-2 ORF3a Monoclonal Antibody (Catalog # MAB107061) at 0.3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to immunoreactive profiles scattered throughout the tissue. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: V
• Applications: WB, IHC, ICC/IF
• Clone: 2819D
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

ORF3a Antibody (2819D) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: SARS-CoV-2 ORF3a peptide
• Specificity: Detects SARS-CoV-2 ORF3a in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-25 ug/mL
  • Immunohistochemistry 0.3-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ORF3a Antibody (2819D) [Unconjugated]

• ORF3a
• U274
• X1

Background

ORF3a is an open reading frame coronavirus protein involved in virus replication in release.  Analysis of ORF3a nucleotide and protein sequences can predict their ability to alter viral cycle and provides insight into the biology of coronaviruses.  ORF3a has been characterized in both SARS-CoV which caused the SARS outbreak and in SARS-CoV-2 which caused the COVID-19 pandemic.  ORF3a is a high priority target that is amenable to drug treatment for COVID-19 post viral syndrome.  Mutations in ORF3a can help understand viral virulence and design suitable therapeutics.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.