Assay Development!

To document assay development for the Zivkovic's lab. We are interested in investigating functional markers of high-density lipoproteins ("good cholesterol") as measure of health and disease status.

Antioxidant Capacity Assay

• This cell-free assay can be performed on a 96-well plate.
• CVs were decent ~1-12%.
• Works well with ApoB-depleted plasma and most likely plasma.
• Not compatible with HDL isolated by UC+FPLC.

Lecithin–Cholesterol Acyltransferase (LCAT) Activity Assay

• This cell-free assay can be performed on a 96-well plate.
• CVs were fantastic <3%.
• Works well with HDL isolated by UC+FPLC, plasma and ApoB-depleted plasma.
• Clear dose reponse with increasing sample concentration.

Cholesterol Efflux Capacity (CEC) Assay

• This cell-based assay is be performed on a 96-well plate.
• New method is robust, most CVs <3%.
• Works with ApoB-depleted plasma and isolated HDL (UC+FPLC). Does not work with untreated plasma (media clumps with plasma).
• Works on multiple cell lines. Cells tested on J774A.1 and HMC3 microglia.