Detects rat Nesfatin-1/Nucleobindin-2 in direct ELISAs and mouse and rat Nesfatin-1/Nucleobindin-2 in Western blots. In direct ELISAs, approximately 7% cross-reactivity with recombinant human Nesfatin-1/Nucleobindin-2 is observed.
Polyclonal Sheep IgG
E. coli-derived recombinant rat Nesfatin-1/Nucleobindin-2 Pro26-Leu106 Accession # Q9JI85
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Detection of Mouse and Rat Nesfatin‑1/Nucleobindin‑2 by Western Blot.
Western blot shows lysates of rat liver tissue and mouse pancreas tissue. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Mouse/Rat Nesfatin‑1/Nucleobindin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6895) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Nesfatin‑1/Nucleobindin‑2 at approximately 50 to 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Nesfatin‑1/Nucleobindin‑2 was detected in immersion fixed frozen sections of rat brain (cortex) using Sheep Anti-Mouse/Rat Nesfatin‑1/Nucleobindin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6895) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to neuronal cell bodies and processes. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Detection of Rat Nesfatin‑1/Nucleobindin‑2 by Simple WesternTM.
Simple Western lane view shows lysates of rat liver tissue, loaded at 0.2 mg/mL. A specific band was detected for Nesfatin‑1/ Nucleobindin‑2 at approximately 55 kDa (as indicated) using 20 µg/mL of Sheep Anti-Mouse/Rat Nesfatin‑1/Nucleobindin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6895) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
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 presumably secreted peptide derived from the translation product of the NUCB2 gene. Nesfatin‑1 is associated with neurons involved in feeding (ARH and PVH), fluid intake (SON and PVH) and autonomic activity, in beta -cells of the pancreas, endocrine cells in the stomach, and in adipocytes. Its presence peripherally has an anorexigenic effect. Mature rat Nesfatin-1 is 82 amino acids (aa) in length. Although it is 10 kDa in MW, its presence in SDS-PAGE is difficult to detect in biological fluids. It represents the N-terminal cleavage product of a 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). Full-length NUCB2 is 48-55 kDa in size, and may be present extracellularly. There is a potential for multiple cleavages that would generate Nesfatin-1, Nesfatin-2 (aa 109-187) and Nesfatin-3 (aa 190-420). A 25 kDa peptide that represents Nesfatin-1 and -2 has been reported. Rat Nesfatin-1 (aa 25-106) shares 97% and 84% aa identity with mouse and human Nesfatin-1, respectively.
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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