Detection of Human Ghrelin/Obestatin by Western Blot.
Western blot shows lysates of human stomach tissue. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human Ghrelin/Obestatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8149) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Ghrelin/Obestatin at approximately 13 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Ghrelin/Obestatin in Mouse Stomach.
Ghrelin/Obestatin was detected in immersion fixed frozen sections of mouse stomach using Sheep Anti-Human Ghrelin/Obestatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8149) at 10 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 is shown at both 60x magnification (upper panel) and 20x magnification (lower panel). Staining was localized to cytoplasm and gastric endocrine cells, respectively. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Ghrelin/Obestatin in Human Stomach.
Ghrelin/Obestatin was detected in immersion fixed paraffin-embedded sections of human stomach using Sheep Anti-Human Ghrelin/Obestatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8149) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of gastric mucosa. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
GHRL (Ghrelin/Obestatin Prepropeptide) is a 117 aa precursor peptide that is processed into three chains, Ghrelin-27 (aa 24-50), Ghrelin-28 (aa24-51) and Obestatin (aa76-98). The prepropeptide is cleaved into proGhrelin, and then further processed into mature Ghrelin by prohormone convertases. Mature Ghrelin is modified with an N-octanoyl group on serine 3 which is required for receptor binding. Ghrelin and Obestatin are predominantly synthesized in the gastric mucosa. Ghrelin plays a role in growth factor release and appetite suppression as well as many other functions in a variety of organs. Obestatin is a putative hormone that has been suggested to have opposite effects on growth factor release and appetite as Ghrelin.
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