24
edits
Changes
m
no edit summary
#To each well, add 80 uL of glycerol reagent. Incubate plate at 37 deg C for 30 minutes.
#Read the plate at 540 nm. This is the initial reading.
#To each well, add 20 uL of triglyceride reagent. Incubate plate at 37 deg C for ~5 minutes. The triglyceride reagent contains a lipase that breaks down the triglycerides into glycerol and fatty acids. The assay measures the glycerol in the sample, so the first reading tells us how much glycerol is present before lipolysis is stimulated. #Re-read the plate at 540 nm. This is the final reading. The second reading tells us how much glycerol is present after all lipolysis has been stimulated and the triglycerides present have been broken down.
#Calculations
##Subtract the initial reading from the final reading. This tells us what proportion of the glycerol in the sample came from triglycerides.
##For each of the initial, final and calculated absorbances, calculate the concentration of the samples from the standard curve.
