pHrodo™ Green E. coli BioParticles™ Phagocytosis Kit for Flow Cytometry

The pHrodo™ Green E. coli 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 of 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.

View a selection guide for all pHrodo Indicators used in imaging, flow cytometry and microplate assays.

The pHrodo™ Green E. coli 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 E. coli 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 E. coli BioParticles™ conjugates are inactivated, unopsonized E. coli 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.

Use with Bacteria
Analysis of phagocytosis by Gram-positive or Gram-negative bacteria is easily accomplished using the ready-made pHrodo™ Green BioParticles™ conjugates. Use ready-made pHrodo™ Green BioParticles™ conjugates for convenient analysis of phagocytosis of Gram-positive or Gram-negative bacteria, or the amine-reactive pHrodo™ Green STP ester for labeling microorganisms or proteins of your choice.

pHrodo™ Indicators for Phagocytosis
The pHrodo™ Green E. coli BioParticles™ Phagocytosis Kit is part of a larger product offering of reagents for measurement of phagocytosis. Scientists can use a variety of pHrodo™ reagents in plate readers, fluorescence microscopy imaging and flow cytometry.

Additional information about pHrodo™ Green products, pHrodo™ Red products, and other pHrodo™ indicators for phagocytosis.