Detection of Human NM23‑H1 by Western Blot. |
Western blot shows lysates of A431 human epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, and K562 human chronic myelogenous leukemia cell line. PVDF Membrane was probed with 0.5 µg/mL of Mouse Anti-Human NM23‑H1 Monoclonal Antibody (Catalog # MAB6256) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, recombinant human NM23-H1 and recombinant human NM23-H2 (10 ng/lane) were included. A specific band was detected for NM23‑H1 at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
NM23-H1 (Non-metastatic protein 23 homolog 1; also NDKA) is a 19‑20 kDa member of the NDK family of enzymes. NM23-H1 is ubiquitous in expression and performs multiple functions. It forms disulfide-linked homohexamers, and heterohexamers with NM23-H2, generating a nucleoside diphosphate kinase that catalyzes a phosphoryl transfer from ATP to a nucleoside diphosphate. It also shows His and Ser/Thr protein kinase activity and forms covalent linkages with molecules diverse as p53 and STRAP. It is found both intracellularly and in blood at ng/mL concentrations. Human NM23-H1 is 152 amino acids (aa) in length, contains one NDP kinase domain (aa 5‑134), and shows acetylation at Ala2 and Lys56, plus phosphorylation at Tyr52, Thr94, Ser122, and Ser125. Human NM23-H1 shares 89% aa identity with human 17‑18 kDa NM23-H2 and 94% aa identity with mouse NM23-H1. A second H1 isoform named NM23-H1B with 25 additional aa at the N‑terminus has also been described.
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