SARS-CoV-2 Spike RBD Antibody

Catalog #: MAB10827 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB10827-100
MAB10827-SP

Detection of SARS-CoV-2 Spike RBD by Western Blot.

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SARS-CoV-2 Spike RBD Antibody Summary

Species Reactivity

SARS-CoV-2

Specificity

Detects SARS-CoV Spike RBD in direct ELISAs.

Source

Monoclonal Mouse IgG_2A Clone # 1042425

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Chinese Hamster Ovary cell line CHO-derived SARS-CoV Spike RBD
Arg306-Phe527
Accession # NP_0828851.1

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 SARS-CoV-2 Spike S1 RBD

Flow Cytometry

0.25 µg/10⁶ cells

SARS-CoV-2 Spike S1 protein bound to ACE-2 in HEK293 Human Cell Line Transfected with Human ACE-2 and eGFP

Immunocytochemistry

3-25 µg/mL

immersion fixed CHO cell line transfected with SARS-CoV-2 Spike

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 SARS-CoV-2 Spike RBD by Western Blot. Western blot shows recombinant SARS-CoV-2 Spike S1 RBD. PVDF membrane was probed with 2 µg/mL of Mouse Anti-SARS-CoV-2 Spike RBD Monoclonal Antibody (Catalog # MAB10827) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Spike RBD at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Flow Cytometry

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Detection of SARS-CoV-2 Spike S1 protein bound to ACE-2 in HEK293 Human Cell Line Transfected with Human ACE-2 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with human ACE-2 and eGFP was incubated with Recombinant SARS-CoV-2 Spike S1 Subunit His-Tag protein (10522-CV), then stained with (A) Mouse Anti-SARS-CoV-2 Spike S1 Monoclonal Antibody (Catalog # MAB10827) or (B) Mouse IgG2A Isotype Control Antibody (MAB003) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B). Staining was performed using our Staining Membrane-associated Proteins protocol.

Immunocytochemistry

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Detection of Spike RBD in CHO Chinese hamster ovary cell line. Spike RBD was detected in immersion fixed CHO Chinese hamster ovary cell line transfected with SARS-CoV-2 (positive staining) and CHO Chinese hamster ovary cell line (non-transfected, negative staining) using Mouse Anti-SARS-CoV-2 Spike RBD Monoclonal Antibody (Catalog # MAB10827) 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

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.

Background: Spike RBD

SARS-CoV was discovered in association with cases of severe acute respiratory syndrome (SARS) that infected more than 8,000 persons with over 900 fatalities worldwide in 2002-2003 (1). It belongs to a family of viruses known as coronaviruses that also include MERS and SARS-Cov2 that causes the global pandemic coronavirus disease 2019 (Covid-19). Coronavirus is commonly comprised of four structural proteins: Spike protein(S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) (1). SARS-CoV S Protein is a type-I trimerized membrane glycoprotein that mediates membrane fusion and viral entry. As with most coronaviruses, proteolytic cleavage of the S protein into two distinct peptides, S1 and S2 subunits, is required for activation. The S1 subunit is focused on attachment of the protein to the host receptor while the S2 subunit is involved with cell fusion (2-4). A metallopeptidase, angiotensin-converting enzyme 2 (ACE-2), has been identified as a functional receptor for SARS-CoV through interaction with a receptor binding domain (RBD) located at the C-terminus of S1 subunit (5, 6). Based on amino acid (aa) sequence homology, the RBD domain of SARS-Cov shares 73% and 24% homology with RBD domain of SARS-CoV2 and MERS, respectively. Before binding to the ACE-2 receptor, structural analysis of the S1 trimer shows that only one of the three RBD domains in the trimeric structure is in the "up" conformation. This is an unstable and transient state that passes between trimeric subunits but is nevertheless an exposed state to be targeted for neutralizing antibody therapy (7). Antibodies to S protein especially the RBD region of SARS-CoV have been shown to inhibit interaction with the ACE-2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (8).

References