Triglyceride Assay from Tissue Culture Cells: Difference between revisions

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 ##Prepare reagent. You need 560ul '''(80ul)''' of glycerol reagent and 140ul '''(20ul)''' of triglyceride reagent. Make extra and combine in a Falcon tube.
 ##Aliquot 700ul into a cuvette or '''100ul into a well of a 96 well plate'''
-##For standards, add 0-5ul of glycerol standard (standard 1-1ul of glycerols standard, 2-2ul, etc.)---If you have reason to believe your signal is low, or if you do not have an idea of the level of signal of your samples you may want to include a second set of diluted standards. In this case you should make a 1:10 dilution of the glycerol standard and water and add 4, 6, 8, and 10ul of that).
+##For standards, add 0-5ul of glycerol standard (standard 1-1ul of glycerol standard, 2-2ul, etc.)---If you have reason to believe your signal is low, or if you do not have an idea of the level of signal of your samples you may want to include a second set of diluted standards. In this case you should make a 1:10 dilution of the glycerol standard and water and add 4, 6, 8, and 10ul of that).
 ##Add 5ul of resuspended lipid to each well to start (also make a 5ul blank of the butanol mixture) and mix.
 ##Let sit for ~30 mins @ room temperature (or 5mins @ 37C if you are in a hurry). If using >10ul of butanol mix the solution may be cloudy. Let it settle and it should become more clear.
 ##Measure absorbance @ 540nm
 ##If any samples are A540<0.1 or above the 5ul standard the A540, then repeat with more or less lipid as required.