Mouse Leptin/OB Antibody

Leptin/OB in Mouse Brain.

Leptin/OB was detected in perfusion fixed frozen sections of mouse brain using Mouse Leptin/OB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF498) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal cytoplasm. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse Leptin/OB by Western Blot

Leptin inhibits adipocyte Tet2 expression via JAK2-STAT3 signaling pathway in adipocytes.aTet2 mRNA levels relative to 36b4 in differentiated mature adipocytes that were treated with PBS, leptin (1 μg/mL), TNF-alpha (5 ng/mL), LPS (500 ng/mL), IFN-gamma (2 ng/mL), or TGF-beta 1 (10 ng/mL) for 24 h (n = 3). b-cTet2 expression in differentiated mature adipocytes treated with leptin at various time points (b) and concentrations (c). Time points and concentrations are indicated on the graphs (n = 3). d Leptin levels in conditioned medium (CM) from iWAT and eWAT of ND-fed and HFD-fed mice (n = 4).eTet2 expression in mature adipocytes treated with control antibody (IgG) or leptin neutralizing antibody (mLeptin) for 1 h, followed by treating with CM from eWAT of ND-fed and HFD-fed mice for another 24 h (n = 3). fTet2 mRNA levels relative to 36b4 in adipocytes of iWAT and eWAT from WT and ob/ob mice (n = 5 WT, n = 6 ob/ob). g Representative immunoblot images of TET2 in adipocytes of iWAT and eWAT from the mice shown in f and densitometry analysis. beta-actin was used as a loading control (n = 3 mice/group). hTet2 expression in differentiated 3T3-L1 adipocytes treated with DMSO (n = 4) or AZD1480 (JAK2 inhibitor, n = 3), Ctrl-siRNA (n = 3) or JAK2-siRNA (n = 3) for 24 h, followed by treating with leptin for another 24 h. iTet2 expression in differentiated 3T3-L1 adipocytes treated with Ctrl-siRNA or STAT3-siRNA for 24 h, followed by treating with leptin for another 24 h (n = 3). All data are presented as mean ± SEM. n indicates the number of biologically independent samples examined. P-values are indicated on the graph. Statistical values are determined by two-sided unpaired Student’s t-test. Source data are provided as a Source Data File. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41467-024-46783-x), licensed under a CC-BY license. Not internally tested by R&D Systems.