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Adiponectin

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Adiponectin is a protein that is involved in regulating glucose and fatty acid metabolism. It is secreted by adipose tissue and plays a role in insulin sensitivity and energy homeostasis.

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Lab products found in correlation

β actin, by Santa Cruz Biotechnology (5 mentions) Interleukin 6 (il 6), by Abcam (4 mentions) Nefa c kit, by Fujifilm (3 mentions) Pparγ, by Cell Signaling Technology (3 mentions) C ebpα, by Cell Signaling Technology (3 mentions) Leptin, by Abcam (3 mentions) Pparγ, by Abcam (3 mentions) Irisin, by Enzo Life Sciences (2 mentions) Myostatin, by Enzo Life Sciences (2 mentions) Pgc 1α, by Cell Signaling Technology (2 mentions) Rat mouse insulin elisa kit, by Merck Group (2 mentions) P acc1, by Cell Signaling Technology (2 mentions) Antibodies against f4 80, by Abcam (2 mentions) Fndc5, by Abcam (2 mentions) Elisa kits for measuring irisin, by Abcam (2 mentions) P smad2, by Cell Signaling Technology (2 mentions) P smad3, by Cell Signaling Technology (2 mentions) β actin, by Abcam (2 mentions) Leptin, by Thermo Fisher Scientific (2 mentions) Interleukin 6 il 6, by Abcam (2 mentions)

Close spelling variants of this product

Mouse adiponectin ELISA kit (13 citations) Anti-adiponectin (12 citations) Human Adiponectin ELISA kit (6 citations) Rat Adiponectin ELISA kit (5 citations) Adiponectin antibody (4 citations) Human adiponectin (4 citations) Adiponectin Human in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit (2 citations) Recombinant human adiponectin (2 citations) Adiponectin receptor 1 (2 citations) Anti-adiponectin antibody (2 citations)

44 protocols using adiponectin

1

Immortalized Renal Proximal Tubule Cell Culture

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Immortalized renal proximal tubule (RPT) cells from WKY rats and SHRs were cultured at 37°C in 95% air and 5% CO2. The RPT cells were grown in DMEM/F12 with insulin, transferrin, epidermal growth factor, and dexamethasone [10 (link),11 (link),16 (link)]. The RPT cells (95% confluence) were serum-deprived for 2 h, and then treated with vehicle, adiponectin (Biovision), Compound C (adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) antagonist, 10−7 mol/l), NG-nitro-l-arginine methyl ester (l-NAME; non-selective NOS inhibitor, 5 × 10−7 mol/l) (Sigma–Aldrich), alone, or in combination. The inhibitors were added 30 min before the addition of adiponectin. Then, the treated cells were extracted in cold cell lysis buffer, sonicated, kept on ice for 1 h, then centrifuged at the speed of 14000 ×g for 30 min to collect the supernatant.
Zhang Y., Wang S., Huang H., Zeng A., Han Y., Zeng C., Zheng S., Ren H., Wang Y., Huang Y., Jose P.A., Ma X.L., Zeng C, & Chen K. (2020). GRK4-mediated adiponectin receptor-1 phosphorylative desensitization as a novel mechanism of reduced renal sodium excretion in hypertension. Clinical science (London, England : 1979), 134(18), 2453-2467.
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2

Molecular Markers of Metabolic Health

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The primary antibodies against p-Akt (Ser473), Akt, p-Smad2, p-Smad3, PPARγ, p-ACC1 and PGC1-α were from Cell Signaling Technology (Beverly, MA). Antibodies against F4/80, CD68, Fndc5 and UCP-1 were from Abcam (Cambridge, MA). ELISA kits for measuring irisin, IL-6 or adiponectin were from BioVision (Milpitas, CA), eBioscience (San Diego, CA) or R&D Systems (Minneapolis, MN) respectively. The recombinant proteins, irisin or myostatin were obtained from Enzo life Sciences (Farmingdale, NY) or R&D systems. The anti-myostatin peptibody (peptibody) was from Atara Biotherapeutics (Westlake Village, CA) (21 (link);22 (link)). Serum insulin concentration was measured using the Rat/Mouse Insulin ELISA kit (Millipore, Billerica, MA). Serum free fatty acid levels were measured using the NEFA C kit from Wako (Richmond, VA).
Dong J., Dong Y., Dong Y., Chen F., Mitch W.E, & Zhang L. (2015). Inhibition of myostatin in mice improves insulin sensitivity via irisin-mediated cross talk between muscle and adipose tissues. International journal of obesity (2005), 40(3), 434-442.
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3

GLP-1 Secretion in GLUTag Cells

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GLUTag cells [31] (link) were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin, and streptomycin. The cells were plated at 50,000 cells/cm2 in 12-well culture plates and allowed to grow for 48 h. The cells were preincubated with KRH buffer (117 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl2, 1.2 mM MgSO4, 20 mM HEPES, and 0.1% BSA) for 30 min. Then, the cells were incubated with 10 μM forskolin (Fsk)/1-methyl-3-(2-methylpropyl)-7H-purine-2,6-dione (IBMX), 100 nM leptin (Sigma), 10 ng/ml resistin (R&D Systems), or 30 μg/ml adiponectin (recombinant mouse gAcrp30; BioVision) in KRH buffer containing 2.5 mM glucose for 2 h at 37°C. The incubation buffer was collected and centrifuged to remove any floating cells. Then, GLP-1 was subjected to ELISA.
Kihira Y., Miyake M., Hirata M., Hoshina Y., Kato K., Shirakawa H., Sakaue H., Yamano N., Izawa-Ishizawa Y., Ishizawa K., Ikeda Y., Tsuchiya K., Tamaki T, & Tomita S. (2014). Deletion of Hypoxia-Inducible Factor-1α in Adipocytes Enhances Glucagon-Like Peptide-1 Secretion and Reduces Adipose Tissue Inflammation. PLoS ONE, 9(4), e93856.
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4

Serum Biomarker Quantification Protocol

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Serum LPS was quantified using a commercial kit based on the guidelines provided by the supplier (Lal limulus amebocyte lysate assay; Cambrex Bio Science, East Rutherford, NJ, USA). Serum free fatty acids (Biovision, Milpitas, CA, USA), adiponectin (Biovision) and leptin (mouse leptin ELISA; Abcam, Cambridge, MA, USA) were determined using commercial detection kits. Serum triglycerides were quantified using a commercial detection kit (Biovision). Serum IL-1β, IL-6 and TNF-α were examined with commercial ELISA kits according to manufacturer’s instructions (R&D Systems, Minneapolis, MN, USA).
Chang C.J., Lu C.C., Lin C.S., Martel J., Ko Y.F., Ojcius D.M., Wu T.R., Tsai Y.H., Yeh T.S., Lu J.J., Lai H.C, & Young J.D. (2017). Antrodia cinnamomea reduces obesity and modulates the gut microbiota in high-fat diet-fed mice. International Journal of Obesity (2005), 42(2), 231-243.
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5

Isolation and Treatment of Colonic Organoids

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Colonic crypts were isolated as we have previously described [35 (link), 36 (link)]. Briefly, crypts were isolated by incubation with 20 mM EDTA, mechanical disruption and centrifugation. Isolated crypts were resuspended in Matrigel at a density of 15 crypts/ul (BD Biosciences, San Jose, CA), plated onto 24-well plates and maintained in complete medium containing advanced Dulbecco’s modified Eagle’s medium/F12, (ADF; Life Technologies, Grand Island, NY), epidermal growth factor (50 ng/ml; Life Technologies), Noggin (100 ng/ml; Peprotech, Rocky Hill, NJ), R- Spondin (500 ng/ml; Sino Biological, Beijing, China), N2 supplement (1.; Invitrogen), B27 supplement (1X; Life Technologies), N-acetylcysteine (1 μM; Sigma, St Louis, MO) and Wnt-conditioned medium as described previously by Barker et al. [37 (link)]. Colonic crypts begin to bud around 2-3 days and on day 3 the organoid cultures from lean and obese Lgr5-GFP mice were treated directly with the recombinant leptin (5-100 ng/ml, BioVision Cat. #4367), adiponectin (5-50 μg/ml, BioVision Cat. #4902), the adiponectin receptor agonist AdipoRon (5-50 uM BioVision Cat. #42565) or dimethyl sulfoxide (DMSO; control vehicle) for 48 hours.
DeClercq V., McMurray D, & Chapkin R. (2015). Obesity promotes colonic stem cell expansion during cancer initiation. Cancer letters, 369(2), 336-343.
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6

Molecular Markers of Metabolic Health

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The primary antibodies against p-Akt (Ser473), Akt, p-Smad2, p-Smad3, PPARγ, p-ACC1 and PGC1-α were from Cell Signaling Technology (Beverly, MA). Antibodies against F4/80, CD68, Fndc5 and UCP-1 were from Abcam (Cambridge, MA). ELISA kits for measuring irisin, IL-6 or adiponectin were from BioVision (Milpitas, CA), eBioscience (San Diego, CA) or R&D Systems (Minneapolis, MN) respectively. The recombinant proteins, irisin or myostatin were obtained from Enzo life Sciences (Farmingdale, NY) or R&D systems. The anti-myostatin peptibody (peptibody) was from Atara Biotherapeutics (Westlake Village, CA) (21 (link);22 (link)). Serum insulin concentration was measured using the Rat/Mouse Insulin ELISA kit (Millipore, Billerica, MA). Serum free fatty acid levels were measured using the NEFA C kit from Wako (Richmond, VA).
Dong J., Dong Y., Dong Y., Chen F., Mitch W.E, & Zhang L. (2015). Inhibition of myostatin in mice improves insulin sensitivity via irisin-mediated cross talk between muscle and adipose tissues. International journal of obesity (2005), 40(3), 434-442.
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7

Bronchoalveolar Lavage Fluid Analysis

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BALF was obtained as previously described (14 (link)). ELISA kits were used to measure serum leptin (Invitrogen, Carlsbad, CA), triacylglycerol (TAG, Pointe Scientific, Canton, MI), insulin (Uppsala, Sweden) and adiponectin (Abcam, Cambridge, UK). Blood glucose was measured via a 6h or 14h fast as described in figure legends. Further, BALF albumin was measured with the Mouse Albumin ELISA Kit (Genway Biotech, Inc., San Diego, CA). Total protein in BALF was measured via standard BCA assay (BCA kit, Sigma-Aldrich, St Louis, MO). BALF cytokines (IL-4, IFN-γ, MCP-1, RANTES, KC, IL-17A, IL-10 and TNF-α) were measured using a BioRad Bio-Plex assay (Hercules, CA) per manufacturer’s instructions. IL-4 was not measureable above the lower limit of the assay.
Milner J.J., Rebeles J., Dhungana S., Stewart D.A., Sumner S.C., Meyers M.H., Mancuso P, & Beck M.A. (2015). Obesity increases mortality and modulates the lung metabolome during pandemic H1N1 influenza virus infection in mice1. Journal of immunology (Baltimore, Md. : 1950), 194(10), 4846-4859.
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8

Adiponectin Signaling Pathway Analysis

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Recombinant human full-length adiponectin was obtained from Biobud (Seongnam, Korea). For Western blotting and immunohistochemical analysis, we used specific antibodies against the following proteins: AdipoR1, AdipoR2, Akt, sterol response element binding protein (SREBP)-1, actin (Santa Cruz Biotechnologies, Santa Cruz, CA, USA), APPL, phospho-AMP activated protein kinase (AMPK), AMPK, phospho-Akt (Cell Signaling, Danvers, MA, USA) and adiponectin (Abcam, Cambridge, UK). Palmitic acid, squalene and compound C were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Jung Y.R., Lee J.H., Sohn K.C., Lee Y., Seo Y.J., Kim C.D., Lee J.H., Hong S.P., Seo S.J., Kim S.J, & Im M. (2017). Adiponectin Signaling Regulates Lipid Production in Human Sebocytes. PLoS ONE, 12(1), e0169824.
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9

Serum Lipid and Inflammatory Markers

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Serum triglyceride (TG) level was measured using the Triglyceride Colorimetric Assay Kit (Cayman Chemical, Ann Arbor, MI), total cholesterol, low density lipoprotein cholesterol (LDL), and high-density lipoprotein cholesterol (HDL) levels were determined using Cholesterol Assay Kit - HDL and LDL/VLDL (Abcam, Cambridge, MA). Serum levels of adiponectin (Abcam) and C-reactive protein (CRP; Abcam) were measured using commercially available rat ELISAs. Assays were conducted as described in the protocol provided by the manufacturer.
Poret J.M., Battle C., Mouton A.J., Gaudet D.A., Souza-Smith F., Gardner J.D., Braymer H.D., Harrison-Bernard L, & Primeaux S.D. (2019). The prevalence of cardio-metabolic risk factors is differentially elevated in obesity-prone Osborne-Mendel and obesity-resistant S5B/PI rats. Life sciences, 223, 95-101.
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10

Western Blot Analysis of Adipogenic Proteins

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Protein was extracted in RIPA Lysis buffer (Invitrogen) together with a protease and phosphatase inhibitor cocktail (Invitrogen) and quantified with a Pierce BCA Protein Assay kit. Extracted protein was mixed with NuPAGE lithium dodecyl sulfate sample buffer (Invitrogen) and reducing agent, heated for 10 min at 85°C, and separated in a precast NuPAGE 4%–12% Bis-Tris polyacrylamide gel (Invitrogen) by SDS-PAGE electrophoresis. Separated proteins were transferred to Amersham Hybond PVDF membranes (GE Healthcare Life Sciences) at 100 V for 2 h. Following transfer, membranes were washed and blocked for 1 h and then probed with primary antibodies preincubated in the blocking agent. Primary antibodies used were: SREBP1 (Novus Biologicals; 1:500), FABP4 (Cell Signaling; 1:400), Adiponectin (Abcam; 1:1,000), C/EBPβ (Cell Signaling; 1:500), and C/EBPα (Cell Signaling; 1:500). After washing and incubation with the secondary antibody, immunoreactivity was visualized by chemiluminescence and densitometry performed with an iBright CL1500 Imaging system (Applied Biosystems).
Mikolajczak A., Sallam N.A., Singh R.D., Scheidl T.B., Walsh E.J., Larion S., Huang C, & Thompson J.A. (2021). Accelerated developmental adipogenesis programs adipose tissue dysfunction and cardiometabolic risk in offspring born to dams with metabolic dysfunction. American Journal of Physiology - Endocrinology and Metabolism, 321(5), E581-E591.
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