Triglyceride Assay from Cells and Tissues: Difference between revisions
updated volumes and lysis procedure |
m Deletion of unnecessary information (1/10 Glycerol Dilution) |
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## Prepare reagent, you need 560 uL '''(80uL)''' of glycerol reagent and 140 uL '''(20uL)''' of triglyceride reagent. Make a bit extra and combine in a falcon tube. | ## Prepare reagent, you need 560 uL '''(80uL)''' of glycerol reagent and 140 uL '''(20uL)''' of triglyceride reagent. Make a bit extra and combine in a falcon tube. | ||
## Aliquot 700 uL into a cuvette or '''100 uL into a well of a 96 well plate'''. | ## Aliquot 700 uL into a cuvette or '''100 uL into a well of a 96 well plate'''. | ||
## For standards add 0-5 uL of glycerol standard | ## For standards add 0-5 uL of glycerol standard. | ||
## Add 5 uL of resuspended lipid to each well to start (also make a 5uL blank of the butanol mixture) and mix. | ## Add 5 uL of resuspended lipid to each well to start (also make a 5uL blank of the butanol mixture) and mix. | ||
## Let sit for ~30 min at room temperature (or 5 min at 37C if you are in a hurry). If using > 10 uL of butanol mixture the solution may be cloudy, let it settle and it should become more clear. | ## Let sit for ~30 min at room temperature (or 5 min at 37C if you are in a hurry). If using > 10 uL of butanol mixture the solution may be cloudy, let it settle and it should become more clear. | ||
## Measure absorbance at 540 nm. | ## Measure absorbance at 540 nm. | ||
## If any samples are A540<0.1 or above the 5uL standard A540 then repeat with more or less lipid as required. | ## If any samples are A540<0.1 or above the 5uL standard A540 then repeat with more or less lipid as required. | ||