Phospholipids are major component of the bilayers of all plasma membranes. A single phospholipid molecule consists of a phosphate group on one (“head”), and two side-by-side chains of fatty acids that make up the "tails”. The phosphate head groups can be modified with organic molecules such as Choline (Cho). Cho-containing phospholipids (Phosphatidylcholines, PC) are critical for structural membrane integrity, cellular metabolism and signaling either as individual molecules or precursors of secondary messengers. Changes in global synthesis of Cho-containing phospholipids are an essential parameter in analysis of cellular response to both, physiological and pathological conditions, environmental stress, or drug treatment. To date, phospholipids biochemistry, cell biology and metabolism remain obscure, due to limited methods for their direct cellular visualisation. BioVision’s EZClick™ Global Phospholipid Synthesis Assay Kit offers a simple and robust method to label and visualise newly synthesised phospholipids in vivo. Based on the metabolic incorporation of the choline analogs directly into their structure, modified phospholipid molecules can be detected with high sensitivity and spatial resolution by click chemistry with azide-containing dyes (Ex/Em= 494/521 nm). This kit enables quantitative analyses of global biosynthesis/turnover of Cho-containing phospholipids in cells. Cells show strong incorporation of Cho analogs into all classes of phospholipids that can be assayed by microplate reader and fluorescence microscope. The kit provides sufficient materials for 100 assays.
Biochemical and cellular assays