The precipitation of proteins and lipid extraction from completely homogenized rat skin tissues - Bertin Instruments

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    The precipitation of proteins and lipid extraction from completely homogenized rat skin tissues

    Context

    In this study, protein precipitation and bioactive lipid extraction from rat skin tissues is reported. The combination Solid Phase Extraction and liquid chromatography tandem mass spectrometry (LCMS/ MS) approach was used to measure concentrations of lipid mediators in these tissues. Rat skin tissues were homogenized using CKMix50_7mL Precellys® Lysing Kit and Precellys® Evolution tissue homogenizer combined with Cryolys® cooling unit. In addition, temperatures of homogenates were measured to investigate if temperature of complete homogenate of rat skin tissues remained below 40°C.

    Results

    Fig. 1 Images of a whole rat hindpaw tissue sample (a),
    and homogenized tissue (b) using Precellys® Evolution.

    Fig. 2 Bioactive lipid mediators of the Arachidonic acid
    cascade were successfully measured, by LC-MS/MS, after
    lipids were extracted from the homogenate by solid phase
    extraction. Concentrations of 5-HETE, 8-HETE and 9-HETE
    were measured to be 1.3 ±0.3, 7.9±0.6 and 1.3 ± 0.4
    ng/mg of tissue, respectively.

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