Immunoblotting Analysis of Adiponectin Receptors

Immunoblotting was conducted as described previously [27 (link), 32 (link)]. The membranes were probed with primary antibodies against adipoR1 (rabbit, 1:1000, Abcam), adipoR2 (rabbit, 1:1000, Abcam), RACK1 (rabbit, 1:1000, Abcam), PKCK2α (rabbit, 1:1000, Cell Signaling Technology), CaMKII (rabbit, 1:1000, Abcam), phospho-CaMKII (mouse, 1:800, Abcam), PKCβ1 (mouse, 1:1000, ThermoFisher Scientific), Cav3.2 (rabbit, 1:800, Alomone) and GAPDH (rabbit, 1:3000, Cell Signaling Technology). Blots were washed and subsequently probed with horseradish peroxidase (HRP)-conjugated goat anti-rabbit or goat anti-mouse IgG secondary antibody (1:5000, Abcam). The immunocomplexes were detected with enhanced chemiluminescence (Merck Millipore). The Chin-X Imager System (Shanghai, China) was used to detect the bands, and NIH ImageJ software was used to quantify the protein band intensities.

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Zhang Y., Wei Y., Zheng T., Tao Y., Sun Y., Jiang D, & Tao J. (2023). Adiponectin receptor 1-mediated stimulation of Cav3.2 channels in trigeminal ganglion neurons induces nociceptive behaviors in mice. The Journal of Headache and Pain, 24(1), 117.

Publication 2023

adipoR1 adipoR2 RACK1 CaMKII phospho-CaMKII PKCβ1 GAPDH enhanced chemiluminescence

Anti igg Antibodies Antibody Camkii Chemiluminescence Chin Gapdh Goat Horseradish peroxidase Membranes Mouse Protein Rabbit

Corresponding Organization :

Other organizations : Soochow University, Second Affiliated Hospital of Soochow University, Helmholtz Zentrum München

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Variable analysis

independent variables

Primary antibodies against adipoR1, adipoR2, RACK1, PKCK2α, CaMKII, phospho-CaMKII, PKCβ1, and Cav3.2

dependent variables

Protein band intensities

control variables

GAPDH as a loading control

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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