HCoV-NL63 Human Coronavirus Spike RBD Antibody

Catalog #: MAB11037 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB11037-100
MAB11037-SP

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HCoV-NL63 Human Coronavirus Spike RBD Antibody Summary

Species Reactivity

HCoV-NL63

Specificity

Detects HCoV-NL63 Spike RBD protein in direct ELISAs.

Source

Monoclonal Mouse IgG_2A Clone # 1044936

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Human embryonic kidney cell HEK293-derived HCoV-NL63 Spike RBD protein
Ala475-Asp634
Accession # YP_003767.1

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

Recombinant HCoV-NL63 S1 RBD

Blockade of Receptor-ligand Interaction

In a functional ELISA binding assay, 0.300-3 μg/mL of this antibody will block 50% of the binding of 250 ng/mL of Recombinant HCo-V-NL63 Spike RBD His-tagged Protein (Catalog # 10605-CV) to immobilized Recombinant Human ACE-2 Fc Chimera Protein (Catalog # 10544-ZN) coated at 1 μg/mL (100 µL/well). At 10 μg/mL, this antibody will block >90% of the binding.

Immunocytochemistry

3-25 µg/mL

immersion fixed HEK293 human embryonic kidney cell line transfected with HCoV-NL63

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Spike RBD by Western Blot. Western blot shows recombinant HCoV-NL63 S1 RBD, recombinant SARS-CoV-2 Spike S1 RBD, recombinant HCoV-HKU1 S1 RBD, recombinant HCoV-229E S1 RBD, and recombinant HCoV-OC43 S. PVDF membrane was probed with 2 µg/mL of Mouse Anti-HCoV-NL63 Spike RBD Monoclonal Antibody (Catalog # MAB11037) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Spike RBD at approximately 25 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Blockade of Receptor-ligand Interaction

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HCoV-NL63 Spike RBD Binding to ACE-2 is Blocked by HCoV-NL63 Antibody. In a functional ELISA binding assay, 0.300-3 μg/mL of this antibody will block 50% of the binding of 250 ng/mL of Recombinant HCo-V-NL63 Spike RBD His-tagged Protein (10605-CV) to immobilized Recombinant Human ACE-2 Fc Chimera Protein (10544-ZN) coated at 1 μg/mL (100 µL/well). At 10 μg/mL, this antibody will block >90% of the binding.

Immunocytochemistry

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Detection of HCoV-NL63 Spike RBD in HEK293 Human Embryonic Kidney cell line transfected. HCoV-NL63 Spike RBD was detected in immersion fixed HEK293 human embryonic kidney cell line transfected (positive staining) and HEK293 human embryonic kidney cell line (non-transfected, or HCoV-229E or HCoV-HKU1 transfected, negative staining) using Mouse Anti-HCoV-NL63 Human Coronavirus Spike RBD Monoclonal Antibody (Catalog # MAB11037) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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.

Background: Spike RBD

HCoV-NL63, a virus first isolated from a child suffering from respiratory disease in 2003, 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) (1, 2). Other well-known human coronaviruses include three viruses that cause relatively mild respiratory disease: HCoV-229E, HCoV-HKU1 and HCov-OC43, plus three viruses that cause the Severe Acute Respiratory Syndrome (SARS-CoV), the Middle East Respirator Syndrome (MERS-CoV), and the global pandemic Covid-19 (SARS-CoV2). HCov-NL63 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-NL63 S protein shares high homology (56%) with HCoV-229E, it does not employ CD13 (aminopeptidase N) as the receptor like HCoV-229E. Instead, HCoV-NL63 engages Angiotensin-Converting Enzyme 2 (ACE-2), the same receptor as SARS-CoV and SARS-CoV2, for cellular entry and replication (3). The receptor binding domain (RBD) of HCoV-NL63 is located at C-terminal region of S1 subunit (4, 5). Although NL63-CoV and SARS-CoV do not share structural homology in RBD region, they bind an overlapping region of ACE-2 (6, 7).

References