Triglyceride Assay from Cells and Tissues: Difference between revisions
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##For standards, add 0-5 and .5ul of glycerol standard | ##For standards, add 0-5 and .5ul of glycerol standard | ||
##Add 5ul of resuspended lipid to each well to start (also make a 5ul blank of the butanol mixture) and mix. | ##Add 5ul of resuspended lipid to each well to start (also make a 5ul blank of the butanol mixture) and mix. | ||
Pop any bubbles with tip before incubating. | ##Pop any bubbles with tip before incubating. | ||
##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. | ##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 | ##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. | ##If any samples are A540<0.1 or above the 5ul standard the A540, then repeat with more or less lipid as required. | ||