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Triglyceride Assay from Cells and Tissues

133 bytes added, 18:23, 1 October 2019
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==Materials==
* '''Homogenization Buffer''' (50 mM Tris pH 8, 5 mM EDTA, 30 mM Mannitol, PI inhibitor, can be made in bulk without the PI, PI added fresh) (to make 50 mL of homogenization buffer - 2.5 mL of 1M Tris pH 8, 500 uL of 0.5 M EDTA, 0.273 g Mannitol, fill up the rest with water)* 10M KOH(28.1g in 50 mL of water)
* '''Chloroform/Methanol Mixture''' (2:1)
* '''Butanol Mixture''': 3 mL butanol, 1.66 mL Triton-X114, 0.33 mL Methanol
##Resuspend triglyceride and glycerol reagent with water if necessary
##Calculate how many sample you have (samples + blank + standard curve)
##Prepare reagent. You need 560ul '''(80ul)''' 80uL of glycerol reagent and 140ul '''(20ul)''' 20uL of triglyceride reagent. Make extra and combine in a Falcon tube.
##Aliquot '''100ul into a well of a 96 well plate'''
##For standards, add 0-5 and .5ul of glycerol standard
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