Key Product Details

Species Reactivity

HCoV-229E

Applications

Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2840H
View all Spike RBD Primary Antibodies »

Product Specifications

Immunogen

Human embryonic kidney cell, HEK293-derived HCoV-229E Spike RBD
Ser292-Asp453
Accession # P15423.1

Specificity

Detects both HCoV-229E and HCoV-NL63  in ELISA.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for HCoV-229E Spike RBD Antibody

Detection of Spike RBD by Western Blot.

Western blot shows recombinant SARS-CoV-2 Spike S1 RBD, recombinant HCoV-HKU1 S1 RBD, recombinant HCoV-NL63 S1 RBD, recombinant HCoV-229E S1 RBD, and recombinant HCoV-OC43 S. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-HCoV-229E Spike RBD Monoclonal Antibody (Catalog # MAB11039) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). Specific bands were detected for Spike RBD at approximately 30 and 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Spike RBD in CHO Chinese hamster ovary cell line transfected with HCoV-229E.

Spike RBD was detected in immersion fixed Transfected HCoV-229E CHO cells (positive), Transfected HCoV-NL63 CHO cells (negative), Transfected HCoV-HKU1 CHO cells (negative), Wild Type CHO cells (negative) using Rabbit Anti-HCoV-229E Spike RBD Monoclonal Antibody (Catalog # MAB11039) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for HCoV-229E Spike RBD Antibody

Application
Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed CHO Chinese hamster ovary cell line transfected with HCoV-229E and  HCoV-NL63 S1 RBD

Western Blot

0.1 µg/mL
Sample: Recombinant HCoV-NL63 S1 RBD and recombinant HCoV-229E S1 RBD

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Spike RBD

HCoV-229E belongs to a family of viruses known as coronaviruses 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).  HCoV-229E is a member of the alpha-coronavirus family and was discovered in 1966 (1, 2). Other well-known human coronaviruses include three viruses that cause relatively mild respiratory disease: HCoV-NL63, HCoV-HKU1 and HCov-OC43, plus three viruses that caused the Severe Acute Respiratory Syndrome (SARS-CoV), the Middle East Respirator Syndrome (MERS-CoV), and the global pandemic Covid-19 (SARS-CoV2). HCov-229E 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.  Although HCoV-229E S protein shares high homology (56%) with HCoV-NL63, it does not employ Angiotensin-Converting Enzyme 2 (ACE2) as the receptor like HCoV-NL63. Instead, HCoV-229E engages CD13 (aminopeptidase N) for cellular entry and replication (3). The receptor binding domain (RBD) of HCoV-229E is solely responsible for receptor binding through three extended receptor binding loops (4).

References

  1. Hamre, D. and J.J. Procknow (1966) Proc. Soc. Exp. Biol. Med. 121:190.
  2. Van der Hoek, L. et al. (2004) Nat. Med. 10:368.
  3. Yeager, C.L. et al. (1992) Nature 357:420.
  4. Wong, A.H.M. et al. (2017) Nat. Commun. 8:1735.

Long Name

Spike Receptor Binding Domain

Entrez Gene IDs

3200426 (HCoV-HKU1); 14254594 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)

Gene Symbol

S

UniProt

P15423.1

Additional Spike RBD Products

Product Documents for HCoV-229E Spike RBD Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for HCoV-229E Spike RBD Antibody

For research use only

Related Research Areas

Infectious Diseases

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Protocols

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