Digitonin (C15-1203-989)

General description

Digitonin, a steroidal glycoside derived from the purple foxglove plant (Digitalis purpurea), stands as a versatile non-ionic detergent with diverse applications in cell biology and biochemical research. Its unique properties position it as a valuable tool for investigating cellular membranes, membrane proteins, and intracellular components. Digitonin demonstrates exceptional membrane solubilization capabilities, effectively extracting membrane-bound proteins while preserving their native conformation for studying interactions and activities. Additionally, it can also selectively permeabilize the cholesterol-rich plasma membrane, allowing for the isolation and purification of specific organelles, and facilitating the study of their functions.

The detergent′s cellular permeabilization property extends to various cell types, enabling the study of intracellular components and processes by introducing antibodies, enzymes, or other molecules into cells. In immunocytochemistry experiments, Digitonin plays a crucial role by permeabilizing cells, allowing for the labeling and detection of intracellular proteins and structures, offering insights into cellular organization and protein localization. Furthermore, Digitonin facilitates RNA extraction and purification by lysing cells, a critical step for molecular biology applications such as gene expression analysis and RNA sequencing. The detergent′s utility is also evident in immunoprecipitation experiments, where it solubilizes membrane proteins, enabling the isolation of specific protein-antibody complexes for the study of protein interactions and signaling pathways.

Application

Digitonin has been used in a study to investigate how lipopeptide toxins target biological membranes.

Digitonin has been used:

• to incubate isolated mitochondria for removing the mitochondrial outer membrane for mitoplasts preparation
• in the permeabilization of HeLa cells for in vitro import assay
• as a component in HEPES-buffered saline (HBS) to permeate the cells and release the aequorins for fluorescence and bioluminescence imaging

Biochem/physiol Actions

Digitonin plays a key role in permeabilizing cell membranes. It also helps to solubilize membrane components. Digitonin specifically interacts with cholesterol in the membrane, resulting in pore formation. It helps to lyse eukaryotic cells but not bacterial membranes.

Features and Benefits

• Can extract membrane proteins like supercomplexes from the mitochondrial inner membrane
• Solubilizes 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) large unilamellar vesicles (LUVs) gradually at high temperature
• Canorm micelles at a wide range of concentrations