CAR-MA Phagocytosis Assay
Phagocytosis is an early and critical event that triggers host defense against invading pathogens. Phagocytosis involves a series of events, starting with cell surface receptor binding and recognition of particles, and then forming an actin-rich membrane extension around the particles. The determination of phagocytosis is traditionally determined by measuring the phagocytosis of cellular "substrate". A variety of substrates can be used, including red blood cells, E. coli, and zymosan (yeast) particles.
Creative Biolabs has developed a systematic approach to detect phagocytosis. Each of our phagocytosis assays quantifies the phagocytosis of one of these substrates by macrophages or other phagocytic cells. Quantification is performed in 96-well plates on a standard colorimetric ELISA reader. Our phagocytosis assay is designed to help you analyze the function of specialized immune cells called CAR-MA, or to assess the effects of antibodies and vaccines, or to screen for TLR ligands, phagocytosis activators, and inhibitors.
Phagocytosis Assay Substrates
E. coli Substrate
Macrophages are seeded overnight in 96-well plates. Cytochalasin is used to pretreat macrophages for 1 hour at 37 °C. E. coli particles are then added for 4 hours and then the phagocytic zymosan particles are detected at 450 nm on a plate reader.
Red Blood Cell Substrate
After treating by IgG, red blood cells are incubated with phagocytic cells. Remove non-phagocytic cells. The phagocytic cells are lysed and detected in a 630 nm colorimetric plate reader using a unique proprietary red blood cell substrate.
Zymosan Substrate
The phagocytic cells are incubated with pre-labeled zymosan granules. Non-phagocytic zymosan particles are blocked using a blocking reagent. The phagocytic cells are engulfed and the zymosan substrate is added. The phagocytic zymosan particles are then detected at 405 nm on a plate reader.
General Procedure
For more details, please feel free to contact us for project quotations and more detailed information.
For Research Use Only | Not For Clinical Use
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