Preparation of whole-cell lysates, SDS-PAGE, and immunoblotting were performed as previously described (15 (link)). Western blots were probed with anti-adiponectin (Millipore, Billerica, MA), anti-β-actin, anti-CCAAT/enhancer binding protein-α (C/EBP-α), anti-perilipin, and anti-PPARγ (Cell Signaling Technology, Beverly, MA) antibodies. Next, blots were probed with horseradish peroxidase-conjugated goat anti-mouse (adiponectin) or anti-rabbit (all other proteins) secondary antibodies (Bio-Rad, Hercules, CA). Relative protein expression was evaluated by densitometry using ImageJ version 1.47 (National Institutes of Health), with β-actin used to control for total protein recovery. RNA isolation, cDNA synthesis, and quantitative real time-PCR (qRT-PCR) were performed as previously described (16 (link)). Relative gene expression was evaluated by the ΔCt method (17 (link)), with β-actin used to control for total mRNA recovery. Primers were designed using Primer-BLAST (National Center for Biotechnology Information) and obtained from Integrated DNA Technologies (Supplemental Table 1 ).
Anti adiponectin
Manufactured by Merck Group
Anti-adiponectin is a laboratory reagent used in research and analysis. It is a specialized antibody that binds to and detects the protein adiponectin, which is involved in various metabolic processes. The core function of anti-adiponectin is to facilitate the measurement and quantification of adiponectin levels in samples, enabling researchers to study its role in different biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase conjugated goat anti mouse adiponectin, by Bio-Rad (2 mentions)
Anti rabbit all other proteins secondary antibodies, by Bio-Rad (2 mentions)
Anti β actin, by Cell Signaling Technology (2 mentions)
Anti pparγ, by Cell Signaling Technology (2 mentions)
Anti ccaat enhancer binding protein α c ebp α, by Cell Signaling Technology (2 mentions)
Anti perilipin, by Cell Signaling Technology (2 mentions)
Nf κb p65, by Abcam (1 mentions)
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (1 mentions)
β actin, by Fortis Life Sciences (1 mentions)
Lamin b, by Santa Cruz Biotechnology (1 mentions)
3 protocols using anti adiponectin
1
Protein and Gene Expression Analysis
2
Protein and Gene Expression Analysis
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Preparation of whole-cell lysates, SDS-PAGE, and immunoblotting were performed as previously described (15 (link)). Western blots were probed with anti-adiponectin (Millipore, Billerica, MA), anti-β-actin, anti-CCAAT/enhancer binding protein-α (C/EBP-α), anti-perilipin, and anti-PPARγ (Cell Signaling Technology, Beverly, MA) antibodies. Next, blots were probed with horseradish peroxidase-conjugated goat anti-mouse (adiponectin) or anti-rabbit (all other proteins) secondary antibodies (Bio-Rad, Hercules, CA). Relative protein expression was evaluated by densitometry using ImageJ version 1.47 (National Institutes of Health), with β-actin used to control for total protein recovery. RNA isolation, cDNA synthesis, and quantitative real time-PCR (qRT-PCR) were performed as previously described (16 (link)). Relative gene expression was evaluated by the ΔCt method (17 (link)), with β-actin used to control for total mRNA recovery. Primers were designed using Primer-BLAST (National Center for Biotechnology Information) and obtained from Integrated DNA Technologies (Supplemental Table 1 ).
3
Adiponectin Signaling Pathway Analysis
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Western blotting was performed using the following antibodies: anti-adiponectin (Millipore), anti-adiponectin receptor 1 (AdipoR1) and anti-AdipoR2 (Thermo Scientific), anti-NAD-dependent deacetylase sirtuin 1 (SIRT1) (Millipore), anti-acetyl-CoA carboxylase (ACC), anti-phospho-ACC (Ser79), anti-total AMP-mediated protein kinase (AMPK) and anti-phospho-AMPK (Thr172) (Cell Signaling, Danvers, MA), anti-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (IκBα) (SantaCruz, Paso Robles, CA), anti-nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 (Abcam, Cambridge, United Kingdom). Cytosolic and nuclear fractions to determine cytosolic IκBα and nuclear NF-κB p65 protein expression, respectively, were prepared using NE-PER nuclear and cytoplasmic extraction reagents (Pierce Biotechnology, Inc., Rockford, IL). Antibodies against β-actin (Bethyl Laboratories, Inc., Montgomery, TX) and lamin B (SantaCruz) were used as loading controls. Western blot bands were analyzed using the ImageJ version 1.42 software (National Institutes of Health, Bethesda, MD).
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