Protein Abundance Detection Protocol

Protein extractions were performed as previously described^{44 (link),49}. Primary antibodies HAS2 (Santa Cruz Biotechnology, sc-34068) (1:200 dilution), HSL (Santa Cruz Biotechnology, sc-74489) (1:200 dilution), α Tubulin (Santa Cruz Biotechnology, sc-53030) (1:200 dilution), Actin (Sigma #A4700) (1:1000 dilution), Adiponectin (homemade) (1:1000 dilution) were used. Protein abundance was detected using the one of the following secondary antibodies: Goat anti-Mouse IRDye 680RD (Li-cor 926-68070), Goat anti-rabbit IRDye 800CW (Li-cor 925-32211) or Goat anti-Rat DyLight 800 (Thermo Fisher SA5-10024) at 1:10,000 dilutions. Antibody decorated membranes were then visualized on a Li-Cor Odyssey infrared scanner (Li-Cor Bioscience). The scanned data were analyzed using Odyssey Version 3.0 software (Li-Cor Bioscience).

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Zhu Y., Li N., Huang M., Bartels M., Dogné S., Zhao S., Chen X., Crewe C., Straub L., Vishvanath L., Zhang Z., Shao M., Yang Y., Gliniak C.M., Gordillo R., Smith G.I., Holland W.L., Gupta R.K., Dong B., Caron N., Xu Y., Akgul Y., Klein S, & Scherer P.E. (2021). Adipose tissue hyaluronan production improves systemic glucose homeostasis and primes adipocytes for CL 316,243-stimulated lipolysis. Nature Communications, 12, 4829.

Publication 2021

α Tubulin Adiponectin Goat anti-Mouse IRDye 680RD Goat anti-rabbit IRDye 800CW Li-Cor Odyssey infrared scanner Odyssey Version 3.0 software

Actin Adiponectin Antibodies Antibody Dilutions Goat Has2 Irdye 800cw Membranes Mouse Protein Rabbit α tubulin

Corresponding Organization : The University of Texas Southwestern Medical Center

Other organizations : Children's Nutrition Research Center at Baylor College of Medicine, University of Namur, Washington University in St. Louis, University of Utah, Baylor College of Medicine

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Variable analysis

independent variables

Primary antibodies HAS2 (Santa Cruz Biotechnology, sc-34068)
Primary antibodies HSL (Santa Cruz Biotechnology, sc-74489)
Primary antibodies α Tubulin (Santa Cruz Biotechnology, sc-53030)
Primary antibodies Actin (Sigma #A4700)
Primary antibodies Adiponectin (homemade)

dependent variables

Protein abundance

control variables

Protein extractions were performed as previously described

controls

Positive control: None specified
Negative control: None specified

Annotations

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