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== 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'''
[[ Category: Animal Work]]
[[ Category: Assays ]]
[[ Category: Lipids ]]
* 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'''
[[ Category: Animal Work]]
[[ Category: Assays ]]
[[ Category: Lipids ]]
