SARS-CoV-2 ORF8 Antibody

Name: SARS-CoV-2 ORF8 Antibody
Brand: R&D Systems
SKU: MAB10820
Price: 111 USD

R&D Systems, Inc. a Bio-Techne Brand

Catalog # MAB10820

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Key Product Details

Species Reactivity:	SARS-CoV-2
Immunogen:	E. coli-derived human SARS-CoV-2 ORF8 Met16-His121 Accession # YP_009724369
Applications:	Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Western Blot
Label:	Unconjugated
Source:	Monoclonal Mouse IgG_2A Clone # 1041422
Specificity:	Detects human SARS-CoV-2 ORF8 in direct ELISAs.

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Applications for SARS-CoV-2 ORF8 Antibody

	Recommended Concentration	Sample
Western Blot	2 µg/mL	Recombinamt SARS-CoV-2 ORF8 protein
Immunohistochemistry	5-25 µg/mL	Immersion fixed paraffin-embedded sections of SARS-CoV-2 infected human lung
Immunocytochemistry	8-25 µg/mL	Immersion fixed HEK293 human embryonic kidney cell line transfected with SARS-CoV-2

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Examples for SARS-CoV-2 ORF8 Antibody

	Western Blot Detection of SARS-CoV-2 ORF8 by Western Blot. Western blot shows recombinant SARS-CoV-2 ORF8 protein. PVDF membrane was probed with 2 µg/mL of Mouse Anti-SARS-CoV-2 ORF8 Monoclonal Antibody (Catalog # MAB10820) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for ORF8 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
	Immunocytochemistry ORF8 in HEK293 Human Cell Line Transfected with SARS-CoV-2. ORF8 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 Mouse Anti-SARS-CoV-2 ORF8 Monoclonal Antibody (Catalog # MAB10820) 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 cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.
	Immunohistochemistry ORF8 in SARS-CoV-2 infected human lung. ORF8 was detected in immersion fixed paraffin-embedded sections of SARS-CoV-2 infected human lung using Mouse Anti-SARS-CoV-2 ORF8 Monoclonal Antibody (Catalog # MAB10820) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). 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.

Formulation, Preparation and Storage

Purification	Protein A or G purified from hybridoma culture supernatant
Reconstitution	Reconstitute at 0.5 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
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	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.

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Background: ORF8

SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that are commonly comprised of four structural proteins: Spike protein(S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) (1). M protein is the most abundant structural protein in the coronavirus membrane, and it is predicted to span the membrane three times, with a short N-terminal domain outside the viral envelope and a long C-terminal domain inside the virion (2). SARS-CoV-2 M protein is a 222 amino acid (aa) glycoprotein that is composed of an 18 aa N-terminal domain on the viral surface, 3 transmembrane domains, and a 122 aa C-terminal domain inside the viral envelope. SARS-CoV-2 M protein shares 89.14%, 98.6%, 98.2%, and 38.36% aa similarity with SARS-CoV-1, bat SARS-CoV, pangolin SARS-CoV, and MERS-CoV M proteins, respectively (3). The M protein of coronavirus plays an important role in assembly of viral particles by interacting with other structural proteins, especially with the E protein (4, 5). In SARS-CoV-2, M protein, combined with E protein, regulates intracellular trafficking of the S Protein and its unique furin-mediated processing (6).

Item	Value
References	Wu, F. et al. (2020) Nature 579:265. Mousavizadeh, L. and S. Ghasemi (2020) J. Microbiol. Immunol. Infect. doi:10.1016/j.jmii.2020.03.022. Thomas, S. (2020) Pathog. Immun. 5:342. Masters, P.S. (2006) Adv. Virus. Res. 66:193. Corse, E. and C.E. Machamer (2003) Virology 312:25. Boson, B. et al. (2020) bioRxiv doi:10.1101/2020.08.24.260901.
Long Name	ORF8
Entrez Gene IDs	3200424 (HCoV-HKU1); 43740577 (SARS-CoV-2)
Alternate Names	ORF8

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SARS-CoV-2 ORF8 Antibody

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Applications for SARS-CoV-2 ORF8 Antibody

Scientific Data Examples for SARS-CoV-2 ORF8 Antibody

Western Blot

Immunocytochemistry

Immunohistochemistry

Formulation, Preparation and Storage

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Background: ORF8

Citations for SARS-CoV-2 ORF8 Antibody

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