I want to measure TGs, HDLs, LDLs, Cholesterol in mice/rat's liver, how to prepare liver homogenate for this case?
I wonder what will be the outcome of measuring these parameters in liver homogenates!!!
This is to evaluate lipid metabolism in an animal fed with different new chemicals in order to development of new hypocholesterolemic agents. Also they could be compared with serum levels.
Choosing a homogenisation method largely depends on the outcome of the experiment. Check out this forum:
https://www.researchgate.net/post/Whats_the_best_method_to_homogenize_rat_liver_for_enzymatic_assays
I think the following protocol will hold good for your assay:
"Liver tissue preparation for G6Pas and GP assay. The whole liver was quickly removed from mice, and 100mg of wet hepatic tissue was placed into ice-cold 0.25m sucrose solutions. The mixture was homogenized at 4ºC for 1 min and diluted to 2 mL/100 mg wet liver with the sucrose solution. The homogenate was centrifuged at 4ºC 12000×g for 30 min. The supernatant fluid was collected and frozen for enzymatic assay (Nordlie and Arion, 1965, 1966; Wallis et al., 1999).
Make a homogenate , extract the lipid , Bligh & Dyer, resolve the lipids in trtiton x100 2% and use kits to determine tg tchol