Figure 9

Summary of factors that affect lipid-probe interactions. The RHD-based assay has reduced experimental variability compared to the DCF-based assay. This can be partially explained by the following differences between the RHD-based assay and the DCF-based assay a) Use of a single lipid (HDL) in the RHD-based assay versus 2 lipids (HDL and LDL) in the DCF-based assay b) DHR is relatively stable, and oxidizes at a predictable rate when exposed to air compared to DCFH which is more unstable and prone to auto-oxidation c) Formation of DCFH from DCFH-DA can be mediated by esterases that are carried over variably during the lipid purification process, while DHR requires no activation and is not prone to this effect d) Use of HBS buffer rather than PBS/azide e) Lipid-DHR interactions are less affected by changes in pH compared to DCFH-lipid interactions.