Lipid Extraction (click for protocol)

Free Fatty Acids

HDL (micro method)

Triglyceride TG (micro method)

Cholesterol TC (micro method)
Protocol for DCL Total Cholesterol Assay
Reagents needed:
               Diagnostic Chemicals Ltd catalog number 225-28
               100 mg/dl cholesterol standard (eg Point Scientific catalog number C7509-STD)
               Total sample volume is 25 µl
1. Set up standard curve in columns 1 and 2 as follows:

Row     Chol Stnd    Water
            (100mg/dl)
A         0 (blank)     25
B         1                 24
C         2.5              22.5
D         5                 20
E         10               15
F         20                5
G         25                0

2. Pipette 5 µl plasma into duplicate wells. (5 µl for chow diet; 2.5 µl for high-fat diet)
3. Add 20 µl water to samples (or whatever vol. brings it to 25 µl)
4. Add 175 µl DCL reagent to all wells
5. Incubate at 37°C for 15 minutes
6. Read at 505 nm

FPLC (includes TC/TG & profile for fractions)