Mouse/Rat Nesfatin-1/Nucleobindin-2 Antibody

Catalog # Availability Size / Price Qty
AF6895
AF6895-SP
Detection of Mouse and Rat Nesfatin‑1/Nucleobindin‑2 by Western Blot.
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Citations (2)
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Mouse/Rat Nesfatin-1/Nucleobindin-2 Antibody Summary

Species Reactivity
Mouse, Rat
Specificity
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.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant rat Nesfatin-1/Nucleobindin-2
Pro26-Leu106
Accession # Q9JI85
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Simple Western
20 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Mouse and Rat Nesfatin-1/Nucleobindin-2 antibody by Western Blot. View Larger

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.

Immunohistochemistry Nesfatin-1/Nucleobindin-2 antibody in Rat Brain by Immunohistochemistry (IHC-Fr). View Larger

Nesfatin‑1/Nucleobindin‑2 in Rat Brain. 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.

Simple Western Detection of Rat Nesfatin-1/Nucleobindin-2 antibody by Simple Western<SUP>TM</SUP>. View Larger

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.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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Shipping
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.

Background: Nesfatin-1/Nucleobindin-2

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)
Alternate Names
DNA-binding protein NEFA; NEFA; NEFAGastric cancer antigen Zg4; Nesfatin1; Nesfatin-1; NUCB2; nucleobindin 2; Nucleobindin-2; nucleobinding 2

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Citations for Mouse/Rat Nesfatin-1/Nucleobindin-2 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Loss of Nucleobindin-2 Causes Insulin Resistance in Obesity without Impacting Satiety or Adiposity
    Authors: A Ravussin, YH Youm, J Sander, S Ryu, K Nguyen, L Varela, GI Shulman, S Sidorov, TL Horvath, JL Schultze, VD Dixit
    Cell Rep, 2018-07-31;24(5):1085-1092.e6.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  2. Role of Nesfatin-1 in the Reproductive Axis of Male Rat
    Sci Rep, 2016-09-07;6(0):32877.
    Species: Rat
    Sample Types: Whole Tissue
    Applications: IHC-P

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