Detection of Human, Mouse, and Rat NM23-H1/H2 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, A431 human epithelial carcinoma cell line, HT‑2 mouse T cell line, and Rat‑2 rat embryonic fibroblast cell line. PVDF Membrane was probed with 0.2 µg/mL of Human/Mouse/Rat NM23‑H1/H2 Polyclonal Antibody (Catalog # AF6019) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for NM23‑H1/H2 at approximately 17‑20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
NM23‑H1/H2 was detected in immersion fixed MCF‑7 human breast cancer cell line using Sheep Anti-Human/Mouse/Rat NM23‑H1/H2 Polyclonal Antibody (Catalog # AF6019) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
NM23-H1 (Non-metastatic protein 23 homolog 1; also NDKA) is a 17‑20 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 histidine protein 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 (Non-metastatic protein 23 homolog 2; also NDKB), 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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