Isolation of lipids from tissue
Isolation of lipids from tissue
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Homogenise 1 g of tissue for 1 minute with 10 ml methanol and add 20 ml chloroform and homogenise well for 2 minutes.
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Filter the solid and resuspend it in chloroform: methanol of 2:1 v/v 30 ml and homogenise for 3 minutes.
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Filter and wash the solid again with 30 ml of chloroform and finally with 10 ml methanol.
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Collect all the filtrate and measure the volume and add 0.88% potassium chloride at a ratio of one quarter of total volume.
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Mix well and allow settling.
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Separate the upper layer by aspiration and add water methanol mixture 1:1 at ratio of one quarter of volume of lower layer.
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Again separate the upper layer and evaporate and dissolve the residue in known volume of solvent and use for spotting TLC plates.
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Last modified: Monday, 19 March 2012, 6:27 AM