pHrodo® Cell Labeling Kit for Incucyte® Phagocytosis Assays

The pHrodo^® Cell Labeling Kit for Incucyte^® enables the labeling of your choice of target cells with a pH-sensitive fluorophore for use in Incucyte^® Phagocytosis Assays.

pHrodo^® Cell Labeling Dye for Incucyte^® enables labeling of whole cells with a pH-sensitive fluorophore. These cells are then suitable for use in downstream applications such as phagocytosis of cells. The unique pHrodo^®-based system uses the acidic environment of the phagosome to quantify phagocytosis. As pHrodo^® labeled cells residing in the neutral extracellular solution (pH 7.4) are engulfed by phagocytes and enter the acidic phagosome (pH 4.5–5.5), a substantial increase in fluorescence is observed. In the absence of phagocytes, the fluorescence intensity of the labeled cells remains low. This kit enables real-time evaluation of phagocytic regulation induced by pharmacological agents as well as genetic and environmental factors.

Presentation, Storage and Stability

This comprehensive kit comes with one vial of pHrodo^® Cell Labeling Dye for Incucyte^®, one vial of DMSO used to solubilize the dye, one bottle of pHrodo^® Wash Buffer (100 mL) and one bottle of pHrodo^® Labeling Buffer (100 mL). The pHrodo^® Labeling Dye for Incucyte^® is supplied as a lyophilized solid. When stored at -20°C, the dye is stable for at least six months. Once solubilized, the solution should be used as soon as possible or stored at -20°C and used within one month.

Recommended Use

We recommend that pHrodo^® Cell Labeling Dye for Incucyte^® is prepared at a stock concentration of 1 mg/mL in the sterile DMSO provided. The dye may then be diluted for direct addition to cells suspended in pHrodo^® Labeling Buffer for Incucyte^®. Note that the dye will also bind to any primary amines present in proteins or cellular debris, therefore we recommend that:

• a) Cell lines be washed with pHrodo^® Wash Buffer for Incucyte^® to remove cell culture media and serum
• b) Any primary cells (such as neutrophils extracted from whole blood) be free from contamination

Additional protocol details can be found in the Product Guide in the Documents tab.