Detection of Human Nesfatin‑1/Nucleobindin‑2 by Western Blot.
Western blot shows lysates of AGS human gastric adenocarcinoma cell line and human testis tissue. PVDF membrane was probed with 3 µg/mL of Rat Anti-Human Nesfatin‑1/Nucleobindin‑2 Monoclonal Antibody (Catalog # MAB5949) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Nesfatin‑1/ Nucleobindin‑2 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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 Rat Anti-Human Nesfatin‑1/Nucleobindin‑2 Monoclonal Antibody (Catalog # MAB5949) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to Golgi. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Nesfatin‑1/Nucleobindin‑2 in Human Stomach.
Nesfatin‑1/Nucleobindin‑2 was detected in immersion fixed paraffin-embedded sections of human stomach using Rat Anti-Human Nesfatin‑1/ Nucleobindin‑2 Monoclonal Antibody (Catalog # MAB5949) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS017) and counterstained with hematoxylin (blue). Specific staining was localized to glandular epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.5 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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