ANTI-ALPHA DEFENSIN-5 (HD5) ANTIBODY CLONE 8C8 (C005B-450123)

Specificity

Clone 6H8-1 is a rat monoclonal antibody that detects RNA-directed RNA polymerase NSP12 of the SARS-CoV-2.

Immunogen

A linear peptide corresponding to 19 amino acids from the NSP12 region of SARS-CoV-2 Replicase polyprotein 1ab.

Application

Quality Control Testing

Evaluated by Western Blotting with recombinant SARS-CoV-2 RdRp (NSP12) protein.

Western Blotting Analysis: A 1:250 dilution of this antibody detected recombinant SARS-CoV-2 RdRp (NSP12) protein.

Tested Applications

Immunocytochemistry Analysis: A representative lot detected SARS-CoV-2 RdRp (NSP12) in Immunocytochemistry application (Meng, W., et al. (2023). J Med Virol. 95(1):e28246).

Western Blotting Analysis: A representative lot detected SARS-CoV-2 RdRp (NSP12) in Western Blotting application (Meng, W., et al. (2023). J Med Virol. 95(1):e28246).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

Replicase polyprotein 1ab (UniProt: P0DTD1: also known as pp1ab, EC: 2.7.7.48, RNA-directed RNA polymerase nsp12, Replicase polyprotein NSP12;Pol RdRp) is encoded by the rep (also known as 1a-1b) gene (Gene ID: 43740578) in SARS-CoV-2. SARS-CoV-2 is a positive-strand RNA virus that causes severe respiratory syndrome in human. Its replicase polyprotein 1ab is cleaved into 16 different chains. Non-structural protein 12 (NSP12; aa 4393-5324) is the RNA-directed polymerase (RdRp) that is required for viral replication and transcription. NSP12 is shown to possesses low RNA polymerase activity and requires interaction with cofactors NSP7 and NSP8 to promote polymerase activity. Within the NSP12 is a right-hand RdRp domain (aa S367 to F920) and a nidovirus-specific N-terminal extension domain (aa D60 to R249) that adopts a nidovirus RdRp-associated nucleotidyltransferase (NiRAN) structural design. These two domains are connected by an interface domain (aa A250 to R365). The active site RdRp domain is formed by the conserved polymerase motifs A to G in the palm domain. Motif A that is composed of residues 611 to 626 (TPHLMGWDYPKCDRAM), contains the conserved classic divalent-cation-binding site D618, and motif C (aa 753 to 767 (FSMMILSDDAVVCFN)] contains the catalytic residues (aa 759 to 761). RdRp has become an important therapeutic target to treat COVID‐19. Inhibition of NSP12 activity not only blocks viral replication and reduces the systemic viral load but reduces pathogenesis associated with SARS‐CoV‐2 infection. (Ref.: Meng, W., et al. (2023). J. Med. Virol. 95(10; e28246; Machitani, M., et al. (2022). Virol. J. 19; 213; Gao, Y., et al. (2020). Science. 368(6492); 779-782; Hillen, HS., et al. (2020). Nature. 584(7819); 154-156).

Physical form

Purified rat monoclonal antibody IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Reconstitution

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.