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Human Leptin/OB Antibody

R&D Systems, part of Bio-Techne | Catalog # AF398

R&D Systems, part of Bio-Techne
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AF398
AF398-SP

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Neutralization, Western Blot

Cited:

Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

E. coli-derived recombinant human Leptin/OB
Val22-Cys167
Accession # Q6NT58

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human Leptin/OB Antibody

Detection of Recombinant Human, Mouse, and Rat Leptin/OB antibody by Western Blot.

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.
Leptin/OB antibody in Human Stomach by Immunohistochemistry (IHC-P).

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.

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.

Applications for Human Leptin/OB Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample:  Immersion fixed paraffin-embedded sections of human stomach

Western Blot

0.1 µg/mL
Sample: Recombinant Human Leptin/OB (Catalog # 398-LP), Recombinant Mouse Leptin/OB (Catalog # 498-OB) and Recombinant Rat Leptin/OB (Catalog # 598-LP)

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 of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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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.

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.

Alternate Names

LEP, OB

Entrez Gene IDs

3952 (Human); 16846 (Mouse); 25608 (Rat)

Gene Symbol

LEP

UniProt

Additional Leptin/OB Products

Product Documents for Human Leptin/OB Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Leptin/OB Antibody

For research use only

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