Detection of Human Nesfatin‑1/Nucleobindin‑2 by Western Blot.
Western blot shows lysates of TK‑6 human lymphoblast cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human Nesfatin‑1/Nucleobindin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5949) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Nesfatin‑1/ Nucleobindin‑2 between 10 and 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Nesfatin‑1/Nucleobindin‑2 in Human Brain.
Nesfatin‑1/Nucleobindin‑2 was detected in immersion fixed paraffin-embedded sections of human brain (hypothalamus) using Sheep Anti-Human Nesfatin‑1/Nucleobindin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5949) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal cell bodies. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Nesfatin‑1/Nucleobindin‑2 in Human Mesenchymal Stem Cells.
Nesfatin‑1/Nucleobindin‑2 was detected in immersion fixed human mesenchymal stem cells differentiated into adipocytes using Sheep Anti-Human Nesfatin‑1/Nucleobindin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5949) 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 Golgi. View our protocol for Fluorescent ICC Staining of Stem 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.
Nesfatin-1 (NEFA Encoded Satiety and Fat-influencing protein 1) is a secreted, 10 kDa peptide derived from the translation product of the NUCB2 gene. Nesfatin-1 is associated with neurons involved in feeding (ARH and PVH) and fluid intake (SON and PVH), in beta cells of the pancreas, and in adipocytes. Its presence peripherally has an anorexigenic effect. Mature human Nesfatin-1 is 82 amino acids (aa) in length. It is the secreted N-terminus of a larger, intracellular 420 aa precursor termed NEFA/Nucleobindin-2/NUCB2. NUCB2 contains a signal sequence (aa 1-24), Nesfatin-1 (aa 25-106), a DNA-binding site (aa 171-223), and two EF-hand regions (aa 241-276 and 293-328). Intracellular uncleaved NUCB2 is 45-50 kDa in size. There is a potential for multiple cleavages, generating Nesfatin-2 (aa 109-187) and Nesfatin-3 (aa 190-420). Human Nesfatin-1 shares 85% aa identity with mouse Nesfatin-1.
Entrez Gene IDs:
4925 (Human); 53322 (Mouse); 59295 (Rat)
DNA-binding protein NEFA; NEFA; NEFAGastric cancer antigen Zg4; Nesfatin1; Nesfatin-1; NUCB2; nucleobindin 2; Nucleobindin-2; nucleobinding 2
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