Fatty Acid Extraction from Kidney Tissue

Kidney tissue samples were homogenized in ice-cold PBS, and fatty acids were extracted as previously described (52 (link)). Briefly, 50 mg protein kidney homogenates containing 60 pmol internal standard (docosanoic-22,22,22-D₃ acid) were transferred to glass tubes and subjected to complete hydrolysis using acid hydrolysis reagent (CH₃CN/37% HCl, 4:1) followed by extraction of fatty acids using hexane. The isolated fatty acids were analyzed by electrospray ionization mass spectrometry using a Thermo LTQ Orbitrap Velos mass spectrometer, operated using Xcalibur software (Thermo Fisher Scientific).

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Hammoud S., Ivanova A., Osaki Y., Funk S., Yang H., Viquez O., Delgado R., Lu D., Phillips Mignemi M., Tonello J., Colon S., Lantier L., Wasserman D.H., Humphreys B.D., Koenitzer J., Kern J., de Caestecker M., Finkel T., Fogo A., Messias N., Lodhi I.J, & Gewin L.S. (2024). Tubular CPT1A deletion minimally affects aging and chronic kidney injury. JCI Insight, 9(6), e171961.

Publication 2024

Thermo LTQ Orbitrap Velos Xcalibur software

Corresponding Organization :

Other organizations : Washington University in St. Louis, Hypertension Institute, Vanderbilt University Medical Center, Vanderbilt University, University of Pittsburgh

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Variable analysis

independent variables

Kidney tissue samples

dependent variables

Fatty acid composition

control variables

Protein content of kidney homogenates
Amount of internal standard (docosanoic-22,22,22-D3 acid)

controls

Positive control: Not specified
Negative control: Not specified

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