pHrodo™ Green E. coli BioParticles™ Phagocytosis Kit for Flow Cytometry
pHrodo&trade; Green <i>E. coli</i> BioParticles&trade; Phagocytosis Kit for Flow Cytometry
Invitrogen™

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 bloodRead more
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Catalog number P35381
Price (USD)
1,268.00
Each
Add to cart
Price (USD)
1,268.00
Each
Add to cart
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.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence
For Use With (Equipment)Flow Cytometer
Product TypePhagocytosis Kit
Dye TypepHrodo™ Green
Emission509⁄533
FormatTube(s), Slide(s)
Product LinepHrodo™
Quantity1 kit
Shipping ConditionWet Ice
SpeciesE. coli
Unit SizeEach
Contents & Storage
Contains lysis buffer (10 mL), Buffer B (200 mL), wash buffer (200 mL), and pHrodo™ green E. coli BioParticles™ (5 vials, lyophilized product). Store in refrigerator (2–8°C) and protect from light. Do not freeze.

Frequently asked questions (FAQs)

Are the Invitrogen BioParticles products sterile?

While the bacteria have been attenuated with formaldehyde and alcohol desiccation, the BioParticles products are not considered sterile, and we do not recommend incubation of more than 4 hours. This applies to all of our dye-labeled (pHrodo, Alexa Fluor, etc.) and unlabeled BioParticles products.

How should I store the pHrodo BioParticles included within pHrodo BioParticles Phagocytosis kits for flow cytometry?

The pHrodo BioParticles are supplied within the kits in lyophilized form. We recommend storing the pHrodo BioParticles vial(s) at 2-8 degrees C, desiccated and protected from light. Once reconstituted, the pHrodo BioParticles can be stored at 2-8 degrees C for several weeks, as long as sodium azide is added to a final concentration of 2 mM. If no sodium azide is added, the cell suspension needs to be used right away or on the same day to avoid contamination.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What is the type of bond that attaches the dyes to the BioParticles probes?

We use amine-reactive dyes to covalently attach fluorescent dyes to all of our BioParticles probes such as the Escherichia coli (K-12 strain) BioParticles probes, Staphylococcus aureus (Wood strain without protein A) BioParticles, and the Zymosan A (S. cerevisiae) BioParticles probes.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What cellular processes can be analyzed with a flow cytometer?

-Calcium flux: Each of the Oregon Green calcium indicators binds intracellular calcium with increasing affinity, providing a sensitivity range to match many applications. Oregon Green probes emit green fluorescence at resting levels of Ca2+ and increase their fluorescence intensity 14-fold with increasing Ca2+ concentration. The cell-permeant formulation (Cat. No. O6807) can be loaded in cell media and is compatible with flow cytometry.
-Rhodamine-based calcium indicators comprise a range of probes for large or small changes in Ca2+ concentration. They exhibit a 50-fold increase in fluorescence upon calcium binding and offer a range of wavelengths that can be used in conjunction with GFP or green-fluorescent dyes for multiplexing. Rhod-2, AM (Cat. No. R1245MP), in particular, localizes to mitochondria and can be used with flow cytometry.
-Membrane potential: A distinctive feature of the early stages of apoptosis is the disruption of the mitochondria, including changes in membrane and redox potential. We offer a range of products specifically designed to assay mitochondrial membrane potential in live cells by flow cytometry, with minimal disruption of cellular function. The MitoProbe family of mitochondrial stains (Cat. Nos. M34150, M34151, and M34152) provide quick, easy, and reliable flow cytometric detection of the loss of mitochondrial membrane potential that occurs during apoptosis. MitoTracker dyes (Cat. Nos. M7510 and M7512) are membrane potential-dependent probes for staining mitochondria in live cells. The staining pattern of MitoTracker dyes is retained throughout subsequent flow cytometry immunocytochemistry, DNA end labeling, in situ hybridization, or counterstaining steps. The Mitochondrial Permeability Transition Pore Assay (Cat. No. M34153) provides a more direct method of measuring mitochondrial permeability transition pore opening than assays relying on mitochondrial membrane potential alone. The mitochondrial permeability transition pore (MPTP) is a non-specific channel formed by components from the inner and outer mitochondrial membranes, and appears to be involved in the release of mitochondrial components during cell death.
-Phagocytosis: In phagocytosis, cells internalize particulate matter such as microorganisms, and this process is important for immune responses and during the clearance of apoptotic cells. Probes for studying phagocytosis include BioParticles indicators—bacteria and yeast labeled with fluorescent dyes.
-Tracking phagocytosis using a quench/wash-based assay can report on simple uptake, or a pH indicator can be used to monitor stages in the pathway. We have no-wash assays labeled with pHrodo Red or Green (Cat. Nos. A10010, P35361, P35364, P35365, P35366, and P35367) and no-wash assays for whole blood (Cat. Nos. A10025, A10026, P35381, and P35382), all suitable for flow cytometry.
-pH changes: Sensitive pH determinations can be made in a physiological range using either fluorescent intensity or ratiometric measurements. pHrodo dyes (Cat. Nos. P35373 and P35372) provide signal intensity modulation from pH 2 to pH 9 and with a choice of fluorescent wavelengths. Tracking internalization of fluorescent dextran is a routine method for analyzing pH changes in cellular compartments. Dextran conjugates of pHrodo dyes (Cat. Nos. P35368 and P10361) provide the most complete solution by allowing discrimination of vesicles from early endosomes to lysosomes, with no quench or wash required.
-Reactive oxygen species: Cells that are environmentally stressed usually contain greatly increased levels of reactive oxygen species (ROS). CellROX reagents are fluorogenic probes developed for the detection and quantitation of ROS in live cells. These cell-permeant reagents are non-fluorescent or very weakly fluorescent in the reduced state; however, when oxidized, they become brightly fluorescent and remain localized within the cell. We offer CellROX Green (Cat. No. C10492), CellROX Orange (Cat. No. C10493), and CellROX Deep Red (Cat. No. C10491) Assay Kits validated for flow cytometry.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.