The pHrodo™ Green S. aureus BioParticles™ Phagocytosis Kit for Flow Cytometry enables the detection of phagocytic activity in whole blood samples and live cell lines by flow cytometry. The kit includes all the reagents required for assessing particle ingestion and red blood cell lysis. Sufficient reagents are provided in the kit for performing approximately 100 assays using 100 µL of whole blood per assay.
The pHrodo™ Green S. aureus BioParticles™ assay provides sensitive detection without the need for quenching reagents and extra wash steps, helping save time and minimize the uncertainty of whether particle signals derive from internalized particles.
The pHrodo™ Green S. aureus BioParticles™ Phagocytosis Kit is:
• Specific—fluorogenic, pH-sensitive dye for detection of phagocytosis • Rapid and reproducible—no wash steps or quencher dye required • Flexible—multiplex with reagents excited by other laser lines • Compatible—multiplex with standard reagents and CD markers
How It works The pHrodo™ Green S. aureus BioParticles™ conjugates are inactivated, unopsonized S. aureus which function as highly sensitive, fluorogenic particles for the detection of phagocytic ingestion. The unique pHrodo™ dye-based system measures phagocytic activity based on acidification of particles as they are ingested, eliminating the wash and quenching steps that are necessary with nonfluorogenic indicators of bacterial uptake.
The pHrodo™ Green Dye The new Molecular Probes™ proprietary, pH-sensitive, pHrodo™ Green dye is non-fluorescent at neutral pH, but turns bright green upon acidification. Because it is both fluorogenic and pH-sensitive, the pHrodo™ Green dye can be used as a specific sensor of phagocytic events; acidification of the phagosome following phagocytosis is marked by green fluorescence. It is therefore an ideal tool with which to study phagocytosis and its regulation by drugs and/or environmental factors.
The fluorophore is readily excited with the 488 nm argon-ion laser and fluorescence emission can be collected using a standard 530/30 nm BP filter on most flow cytometers, and thus is easily multiplexable with reagents used on other lasers .
Rapid and reproducible Wash steps and quencher dyes are not needed, since the pHrodo™ Green dye is non-fluorescent outside the cell, making the staining protocol simple and fast. The elimination of wash steps and quencher dyes also improves assay reproducibility, particularly in plate reader-based assays.
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For Research Use Only. Not for use in diagnostic procedures.
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