Human Leptin/OB Antibody

(2 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Leptin/OB in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 25% cross-reactivity with recombinant mouse Leptin/OB is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Leptin/OB
    Val22-Cys167
    Accession # Q6NT58
  • Formulation
    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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize Leptin/OB-induced proliferation in the BaF3 mouse pro‑B cell line transfected with human Leptin R. The Neutralization Dose (ND50) is typically 0.5-3.0 µg/mL in the presence of 7.5 ng/mL Recombinant Human Leptin/OB.
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.
Data Examples
Detection of Recombinant Human, Mouse, and Rat Leptin/OB by Western Blot. Western blot shows 100 ng of Recombinant Human Leptin/OB (Catalog # 398-LP), Recombinant Mouse Leptin/OB (Catalog # 498-OB) and Recombinant Rat Leptin/OB (Catalog # 598-LP). PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human Leptin/OB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF398) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Leptin/OB at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
Immunohistochemistry
Leptin/OB in Human Stomach. Leptin/OB was detected in immersion fixed paraffin-embedded sections of human stomach using Goat Anti-Human Leptin/OB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF398) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in gastric glands. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Cell Proliferation Induced by Leptin/OB and Neutralization by Human Leptin/OB Antibody. Recombinant Human Leptin/OB (Catalog # 398-LP) stimulates proliferation in the BaF3 mouse pro‑B cell line transfected with human Leptin R in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human Leptin/OB (7.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human Leptin/OB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF398). The ND50 is typically 0.5-3.0 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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: Leptin/OB

Leptin is a protein product of the mouse obese gene. Mice with mutations in the obese gene that block the sythesis of Leptin have been found to be obese, diabetic and to have reduced activity, metabolism and body temperature. cDNA clones encoding Leptin have been isolated from human, simian, mouse and rat cells. Human Leptin shares approximately 84% sequence identity with the mouse protein. Human Leptin cDNA encodes a 167 amino acid residue protein with a 21 amino acid residue signal sequence that is cleaved to yield the 146 amino acid residue mature protein. The expression of Leptin mRNA has been shown to be restricted to adipose tissue.

A high-affinity receptor for Leptin (OB-R) with homology to gp130 and the G-CSF receptor has been cloned. OB-R mRNA has been shown to be expressed in the choroid plexus and in the hypothalamus. OB-R has also been identified as an isoform of B219, a sequence that is expressed in at least four isoforms in very primitive hematopoietic cell populations and in a variety of lymphohematopoietic cell lines (1 - 3). The roles of leptin in body weight regulation, hematopoiesis and reproduction continue to be investigated.

  • References:
    1. Considine, R. and J. Caro (1996) Clinical Chemistry 42:843.
    2. Tartaglia, L.A. et al. (1995) Cell 83:1263.
    3. Cioffi, J.A. et al. (1996) Nature Medicine 2:585.
  • Entrez Gene IDs:
    3952 (Human); 16846 (Mouse); 25608 (Rat)
  • Alternate Names:
    FLJ94114; LEP; leptin (murine obesity homolog); leptin (obesity homolog, mouse); Leptin; OB; Obese protein; obese, mouse, homolog of; Obesity factor; OBOBS
Related Research Areas
Citations:

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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Species
Applications
Sample Type
  1. UV-induced inhibition of adipokine production in subcutaneous fat aggravates dermal matrix degradation in human skin
    Sci Rep, 2016;6(0):25616.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Fresh
  2. Increased leptin expression in endometriosis cells is associated with endometrial stromal cell proliferation and leptin gene up-regulation.
    Authors: Wu MH, Chuang PC, Chen HM
    Mol. Hum. Reprod., 2002;8(5):456-64.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: WB
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Normal Goat IgG Control

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