The concentration of FFAs in the adipocyte-CM was quantified using an ELISA assay kit (ab65341; Abcam). Plasma adipokine levels in WT and BaKO mice were measured using the Mouse Adipokine Array Kit (ARY013; R&D Systems) and LEGENDplex (740929; Biolegend). Blood was collected from the retroorbital sinus using a capillary tube.
Mouse adipokine array kit
Manufactured by R&D Systems
Sourced in United Kingdom
The Mouse Adipokine Array kit is a multiplex assay designed to simultaneously detect the relative expression of multiple adipokine proteins in mouse samples. It provides a quick and efficient method for profiling the secreted adipokine levels in various mouse models and experimental conditions.
Automatically generated - may contain errors
5 protocols using mouse adipokine array kit
1
Quantifying Adipocyte-Secreted FFAs
2
Adipokine Profiling in Drp1 Mice
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Adipokines were measured from 100 μL HFD-fed WT and Drp1 KI plasma samples by using the Mouse Adipokine Array kit (ARY013, R&D systems), following the protocol provided by the supplier.
3
Measuring Circulating Hormones and Peptides
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Circulating hormones and peptides levels were assessed from 100μl of plasma samples with Mouse Adipokine Array kit (ARY013, R&D systems), according to manufacturer's instructions. The intensity of the blot was analyzed using the Protein Array analyzer plugin for ImageJ. Active GLP-1 levels in plasma were measured using a U-plex mouse active GLP-1 assay (Ref: K15305K, from Meso Scale Diagnostics, LLC).
4
Western Blot and Adipokine Analysis of Adipogenesis
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Cells and tissues were homogenized into lysis buffer containing 2% sodium dodecyl sulfate (SDS) and 60 mM Tris·HCl (pH6.8). Lysates were quantitated and equal amounts of protein were loaded to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Antibodies used are C/EBPα and HSP90 (Santa Cruz Biotechnology), PPARγ and Pref-1 (Cell Signaling Technology).
For the adipokine array, cells supernatants were analyzed by Mouse Adipokine Array kit (R&D), and quantified by ImageJ.
For the adipokine array, cells supernatants were analyzed by Mouse Adipokine Array kit (R&D), and quantified by ImageJ.
5
Mouse Adipokine Profiling in GL261 Cells
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Determination of relative levels of pre-selected mouse adipokines in conditioned and non-conditioned GL261 cell cultures was performed by using Mouse Adipokine Array Kit (R&D Systems Europe, UK). Cells were rinsed with PBS before Lysis Buffer solubilization at, approximately, 1 × 107 cells/mL. Then, they were resuspended and rocked at 4 °C for 30 min, following a microcentrifugation at 14,000 x g for 5 min. After supernatant transfer into a clean test tube, proteins were quantified a total protein assay. Array Procedure was performed according to manufacture instructions (Catalogue Number ARY024). Data analysis was based on pixel densities on developed X-ray film collected and analyzed using Gel-Doc XR (Bio-Rad© Laboratories), acquired by Image Lab Software 5.2.1 (Bio-Rad© Laboratories) then quantified by ImageJ 1.49 (NIH, USA).
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