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.
Nesfatin-1/Nucleobindin-2 Antibody
R&D Systems | Catalog # AF6895
Scientific images may be resized, cropped, or adjusted for brightness or contrast for presentation purposes. These updates are limited to presentation and do not affect the underlying scientific interpretation of the data. Where available, additional source data may be provided upon request.
Key Product Details
Species Reactivity
Validated:
Mouse, Rat
Cited:
Mouse, Rat
Applications
Validated:
Immunohistochemistry, Western Blot, Simple Western
Cited:
Immunohistochemistry, Immunohistochemistry-Paraffin, Immunocytochemistry
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
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Product Specifications
Immunogen
E. coli-derived recombinant rat Nesfatin-1/Nucleobindin-2
Pro26-Leu106
Accession # Q9JI85
Pro26-Leu106
Accession # Q9JI85
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.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for Nesfatin-1/Nucleobindin-2 Antibody
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 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.
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.
Detection of Nesfatin-1/Nucleobindin-2 by Immunocytochemistry/ Immunofluorescence
EA alleviates surgical trauma-induced overexpression of Nesfatin-1 in the hypothalamus. A Immunofluorescence staining for Nesfatin-1 and c-Fos in the PVN at 24 h post-surgery in the Intact, HT, and EA + HT groups. Scale bar = 100 μm. B, C Quantification of c-Fos-positive cells and their co-labeled with Nesfatin-1 in each group at 24 h post-surgery. D, E Expression levels of Nesfatin-1 mRNA and protein in the hypothalamus in each group. F Immunofluorescence staining for Nesfatin-1 and CRH in the PVN at 24 h post-surgery. Scale bar = 100 μm. G–I Quantification of CRH-positive cells, Nesfatin-1-positive cells, and their co-labeled cells in each group. All data are shown as mean ± SEM, n = 3–6 in each group, * p < 0.05, ** p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://cmjournal.biomedcentral.com/articles/10.1186/s13020-024-00974-2), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Nesfatin-1/Nucleobindin-2 by Western Blot
EA alleviates surgical trauma-induced overexpression of Nesfatin-1 in the hypothalamus. A Immunofluorescence staining for Nesfatin-1 and c-Fos in the PVN at 24 h post-surgery in the Intact, HT, and EA + HT groups. Scale bar = 100 μm. B, C Quantification of c-Fos-positive cells and their co-labeled with Nesfatin-1 in each group at 24 h post-surgery. D, E Expression levels of Nesfatin-1 mRNA and protein in the hypothalamus in each group. F Immunofluorescence staining for Nesfatin-1 and CRH in the PVN at 24 h post-surgery. Scale bar = 100 μm. G–I Quantification of CRH-positive cells, Nesfatin-1-positive cells, and their co-labeled cells in each group. All data are shown as mean ± SEM, n = 3–6 in each group, * p < 0.05, ** p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://cmjournal.biomedcentral.com/articles/10.1186/s13020-024-00974-2), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Nesfatin-1/Nucleobindin-2 by Immunocytochemistry/ Immunofluorescence
EA alleviates surgical trauma-induced overexpression of Nesfatin-1 in the hypothalamus. A Immunofluorescence staining for Nesfatin-1 and c-Fos in the PVN at 24 h post-surgery in the Intact, HT, and EA + HT groups. Scale bar = 100 μm. B, C Quantification of c-Fos-positive cells and their co-labeled with Nesfatin-1 in each group at 24 h post-surgery. D, E Expression levels of Nesfatin-1 mRNA and protein in the hypothalamus in each group. F Immunofluorescence staining for Nesfatin-1 and CRH in the PVN at 24 h post-surgery. Scale bar = 100 μm. G–I Quantification of CRH-positive cells, Nesfatin-1-positive cells, and their co-labeled cells in each group. All data are shown as mean ± SEM, n = 3–6 in each group, * p < 0.05, ** p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://cmjournal.biomedcentral.com/articles/10.1186/s13020-024-00974-2), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Nesfatin-1/Nucleobindin-2 by Western Blot
Knockdown of Nesfatin-1 in PVN alleviates HPA axis hyperactivity and anxiety-like behaviors induced by surgical trauma. A, B Diagram illustrating the sites of virus injection. Scale bar = 50 μm. C, D Expression levels of Nesfatin-1 mRNA and protein in the hypothalamus from different groups. E, F Serum levels of ACTH and CORT in different groups. G, H Expression levels of CRH mRNA and protein in the hypothalamus from different groups. OFT (I–K), EPM (L, M), and LDBT (N, O) were used to assess anxiety levels in different groups of mice. All data are shown as mean ± SEM, n = 4–8 in each group, * p < 0.05, ** p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://cmjournal.biomedcentral.com/articles/10.1186/s13020-024-00974-2), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Nesfatin-1/Nucleobindin-2 Antibody
Application
Recommended Usage
Immunohistochemistry
5-15 µg/mL
Sample: Immersion fixed frozen sections of rat brain (cortex)
Sample: Immersion fixed frozen sections of rat brain (cortex)
Simple Western
20 µg/mL
Sample: Rat liver tissue
Sample: Rat liver tissue
Western Blot
2 µg/mL
Sample: Rat liver tissue and mouse pancreas tissue
Sample: Rat liver tissue and mouse pancreas tissue
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.
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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.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.
Calculators
Background: Nesfatin-1/Nucleobindin-2
Alternate Names
NEFA, Nesfatin1, NUCB2, Nucleobindin-2
Gene Symbol
NUCB2
UniProt
Additional Nesfatin-1/Nucleobindin-2 Products
Product Documents for Nesfatin-1/Nucleobindin-2 Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Nesfatin-1/Nucleobindin-2 Antibody
For research use only
Related Research Areas
Citations for Nesfatin-1/Nucleobindin-2 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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