I am currently working on establishing a diabetes model by inducing insulin resistance using palmitic acid (PA). To prepare the PA, I dissolved a calculated amount in absolute ethanol to create a 100 mM stock solution, which I then heated at 70°C for 10 minutes. Meanwhile, I prepared a 10% BSA solution in low-glucose DMEM with 10% FBS and incubated it in a water bath at 37°C for about 20 minutes and filtered it.

I read that the fatty acid to BSA ratio of 5:1 is considered optimal. Following this, I added 1 ml of 10% BSA to 5 ml of the PA solution. However, after adding the BSA, I noticed the formation of a milky suspension rather than a clear solution, which did not dissolve properly in the treatment media.

Could anyone advise whether my method is correct or suggest a better approach for preparing the palmitic acid solution?