Spike RBD Antibody (1045307) [Unconjugated]

Spike RBD was detected in immersion fixed CHO Chinese hamster ovary cell line transfected with HCoV-HKU1 (positive staining), HCoV-229E transfected (negative staining), HCoV-NL63 transfected cell (negative staining), and CHO Chinese hamster ovary cell line (non-transfected, negative staining) using Mouse Anti-HCoV-HKU1 Spike RBD Monoclonal Antibody (Catalog # MAB10936) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cell cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: V
• Applications: ICC/IF
• Clone: 1045307
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Spike RBD Antibody (1045307) [Unconjugated] Summary

• Immunogen: Human embryonic kidney cell HEK293-derived HCoV-HKU1 Spike RBD
  Thr310-Tyr624
  Accession # Q5MQD0.1
• Specificity: Detects Human Coronavirus HCoV-HKU1 Spike RBD and 229E in direct ELISAs. No staining of NL63 HKu1.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 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 Spike RBD Antibody (1045307) [Unconjugated]

• Spike RBD

Background

HCoV-HKU1 was identified in Hong Kong in 2005 as a new human coronavirus (1). Coronaviruses are a family of viruses that are commonly comprised of a large plus-strand RNA genome and four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein(N). There are two well-known human coronavirus families that infect humans: Alpha coronaviruses which includes HCoV-229E and HCoV-NL63; beta coronaviruses that includes HCov-OC43, Severe Acute Respiratory Syndrome (SARS-CoV), Middle East Respirator Syndrome (MERS-CoV), and global pandemic Covid-19 (SARS-CoV2) (2). The HCoV-HKU1 Spike Protein (S Protein) is a glycoprotein that mediates membrane fusion and viral entry. As with most coronaviruses, proteolytic cleavage of the S protein generates two distinct peptides, S1 and S2 subunits. The S1 subunit is focused on attachment of the protein to the host receptor, while the S2 subunit is involved with cell fusion. The receptor binding domain (RBD) of HCoV-HKU1 is located at C‑terminal region of S1 subunit, similar to SARS‑COV, MERS‑COV and SARS‑COV2, but the RBD regions do not share significant amino acid sequence identity (3). HCoV‑HKU1 has been demonstrated to bind specifically to 9‑O‑acetylated sialic acids (9-O-Ac-Sias) attached as terminal residues to glycan chains on glycoproteins and lipids, but additional receptors remain unknown (4). HCoV‑HKU1, along with HCov-OC43, differ from other cornonaviruses in that their virions possess two types of surface projections, both involved in attachment: large "spikes" that are comprised of homotrimers of the S protein, and unique, smaller protrusions, comprised of the homodimeric hemagglutinin‑esterase (HE) (5).

1. Woo, P. et al. (2005) J. Virol.79:884.
2. Ogimi, C. et al. (2020) J Pediatric Infect Dis Soc doi: 10.1093/jpids/piaa037.
3. Qian, Z. et al. (2015) J. Virol. 89:8816.
4. Huang, X. et al. (2015) J Virol 89:7202.
5. Hulswit, R.J.G. et al. (2019) PNAS 116:2681.

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.

Bioinformatics

• Uniprot:
  • Q5MQD0.1()