Hi everyone,

I am following a protocol to conjugate palmitic and oleic acid with fatty acids-free BSA. I dissolve the acids in 0.1 M NaOH to create a 100mM stock and then I add this to 5% BSA to create a 2.5mM stock. After this, I mix the fatty acids-BSA complex with my media to get a final concentration of 0.350uM of fatty acids. However, when I add this mix to my cells (adipocytes) sometimes I see that the cells break down immediately and when I look through the normal microscope after I change the media I only see lipid droplets floating instead of cells at some parts of the wells. The cells at these parts are gone, so I assume that they break down. Has anyone experienced something like this? This has also happened to my control which has only BSA with NaOH without the fatty acids. Can it be the PH? I don't experience this all the time and now in all the wells of the same plate so it is very confusing.