Detection method and application for high-abundance and low-abundance class phospholipid compounds in cells
A technology for phospholipid compounds and detection methods, applied in the fields of biological sample materials and analysis, to achieve the effects of improving the detection rate, expanding phospholipid omics information, and avoiding interference
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[0078] Detection of high and low abundance phospholipids in RAW264.7 cells
[0079] (1) Pretreatment of RAW264.7 cells
[0080] Take the cells growing in the logarithmic phase, discard the medium, wash the cells twice with PBS, add 1.5 mL of PBS solution pre-cooled at 4°C, blow and beat the cells several times, prepare the cell suspension and count. Take 1 mL of cell suspension in a 1.5 mL centrifuge tube, centrifuge at low temperature, discard the supernatant, and add 40 μL of internal standard solution (PC 17:0-17:0, Lyso PC 17:0, PA 17:0-17:0, LPA17:0, PS 17:0-17:0, LPS 17:1, LPI 17:1, PG 15:0-15:0, PE 17:0-17:0, LPE 17:1) and 1.2mL0. 1N hydrochloric acid aqueous solution / methanol / chloroform mixed solution (1 / 1 / 1, v / v / v), vortex at 2500r for 2 minutes.
[0081] The preparation method of the 0.1N hydrochloric acid aqueous solution is to add 0.9mL concentrated hydrochloric acid into a 100mL volumetric flask, and dilute to the mark with water.
[0082]After standing at 4°C ...
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