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After evaporation of iodine, TAG bands were scraped from the TLC plates and transfer to a Pasteur pipet containing glass wool.
The radioactive bands were scraped from the gel and counted in a liquid scintillation counter (LS 6000 IC; Beckman Coulter Inc., Palo Alto, CA) using UltimaGold™ (PerkinElmer Life And Analytical Sciences, Inc.; Wellesley, MA, USA).
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The DAG band was scraped from the plate and methylated using a mixture of toluene-methanol (BF3/methanol 14%) (20%-55%-25% by volume), as described by Morrison and Smith [24].
Phospholipids, triglyceride, and cholesterol ester bands were scraped into separate tubes, and lipids were extracted from silica with chloroform-methanol and converted into fatty acid methyl esters by boron trifluoride catalysis [ 100].
Iodine stains corresponding to CE bands were scraped and counted.
TG-specific bands were scraped and the radioactivity was measured by liquid scintillation counting.
Comigrating bands were scraped off, extracted with CHCl3/MeOH and dried under N2.
DG bands were scraped, lipids were extracted and dissolved in 100 μl ddH2O containing 1% Triton-X100.
The bands were scraped off, washed with EtOAc/MeOH (1 1) and filtered to yield four compounds (1 4).
After developing with a solvent of 70 20 1 hexane/ethyl ether/acetic acid, the bands were scraped into 13-ml glass vials and esterified using 5%% methanolic HCl.
Individual bands of lipid were scraped from the plate into columns made from Pasteur pipettes that were fitted with 0.2 µm, Type FG Fluoropore membrane filters (Millipore).
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