Description
• Fully quantify phagocytosis with no manual cell counting
• High-throughput 96-well format
• Convenient quantitation in a standard microplate reader
Phagocytosis can be assayed by measuring the engulfment of a cell "substrate". However, traditional assays require tedious cell counting under a microscope. Our CytoSelect 96-Well Phagocytosis Assay, E. coli Substrate provides a more accurate, user-friendly, high-throughput alternative to the standard phagocytosis assay. The assay may be adapted for use with 24-well or 48-well plates.
Introduction: In mammals, phagocytosis by phagocytes (e.g.macrophages, dendritic cells, and neutrophils) is essential for a variety of biological events, including tissue remodeling and the continuous clearance of dying cells. Furthermore, phagocytosis represents an early and crucial event in triggering host defenses against invading pathogens. Phagocytosis comprises a series of events, starting with the binding and recognition of particles by cell surface receptors, followed by the formation of actin-rich membrane extensions around the particle. Fusion of the membrane extensions results in phagosome formation, which precedes phagosome maturation into a phagolysosome. Pathogens inside the phagolysosome are destroyed by lowered pH, hydrolysis, and radical attack (Figure 1). These early events that are mediated by the innate immune system are critical for host survival. As a result of this process, pathogen-derived molecules can be presented at the cell surface (antigen presentation), allowing the induction of acquired immunity.
MyBioSource's CytoSelect 96-well Phagocytosis Assay (E. coli) uses enzyme-labeled, E. coli particles as a phagocytosis pathogen; however, it does not involve subjective manual counting of E. coli particles inside cells. Instead external particles are blocked before the colorimetric detection of engulfed E. coli particles (Figure 2). This format provides a quantitative, high-throughput method to accurately measure phagocytosis. The CytoSelect 96-well Phagocytosis Assay (E. coli) provides a robust system for screening TLR ligands, phagocytosis activators or inhibitors. Each kit provides sufficient quantities to perform 96, 48, 24 tests in a 96, 48, 24-well plate, respectively