Triglyceride and Glycerol Assay in Milk and Serum

Materials

• Sigma Triglyceride Assay Kit (Cat TR0100). Resuspend both the triglyceride (10 mL) and glycerol (40 mL) with water. Reagents are good at 4C for about a month
• Glycerol Standard (0.26 mg/mL or 2.82 mM)
• Serum or milk

Procedure

• Plan out plate, including a blank, samples, and space for 7 standards.
• Add 80 uL of glycerol reagent to each well.
• Add 2.5 uL of sample to each well
• Add 0, 0.5, 1, 2, 3, 4, 5 uL to each of the standard wells.
• Incubate at room temperature for 30 mins (or 5 mins at 37C if you are in a hurry)
• Read plate at 510 nM and 700 nM (for scatter)
• Add 20 uL of triglyceride reagent and wait another 30 mins at room temperature
• Re-read plate at same measures.

Calculations

• Use the standard curve to calculate glycerol levels (in mM) for before and after the triglyceride reagent.
• Glycerol levels are the first value
• Triglyceride levels are the mM Glycerol After - Before.
• To convert to mg/dL use the equation Triglycerides (mg/dL) = mM × 88.57396 g/mol