Phospholipase C activity measurement - (Apr/23/2012 )

Hi,

I'm not sure which section to post this in entirely but Biochemistry seems to be the most logical.

I want to try and measure the activity of Phospholipase C in human endothelial cells and was wondering if anyone had any reliable techniques for measuring it.

I have looked at several papers online and the methods I have found appear to fall into immunoprecipitation followed by phospho-tyrosine immunoblotting, or measurement of enzyme activity with PIP3 or something. Does anyone know which method is more reliable or easier to reproduce?

I was also wondering if it was possible to do colocalisation studies under confocal microscopy.

worthington has a titrimetric assay procedure at this webpage: phospholipase c assay.

some people where i work assay(ed) with pip3 but i don't know the full procedure (it may be radiolabelled, i'm not sure, it's been a long time since i discussed this with them).