Invitrogen™ pHrodo™ Green E. coli BioParticles™ Conjugate for Phagocytosis

Catalog No.	Quantity
P35366	5 x 2 mg

Catalog No. P35366 Supplier Invitrogen™ Supplier No. P35366

Description

Achieve faster staining and more accurate results without the need for wash steps or quencher dye with pH-sensitive pHrodo Green E. coli BioParticles Conjugate for Phagocytosis.

Achieve faster staining and more accurate results without the need for wash steps or quencher dye with pH-sensitive pHrodo Green E. coli BioParticles Conjugate for Phagocytosis. This dye-bioparticle conjugate provides faster and more accurate results than any other dye-bioparticle conjugate and has reduced signal variability and improved timing in sensitive experiments. pHrodo Green dye conjugates are non-fluorescent outside the cell, but fluoresce bright green in phagosomes, specifically detecting intracellular phagocytosis and endocytosis. The pHrodo Green E. coli BioParticles Conjugate can also be multiplexed with a wide variety of blue, red, and far-red dyes reporters such as Mitosox Red, CellEvent Caspase 3/7 Red, NucBlue, RFPs, and Mitotracker Deep Red, among many others.

The fluorescence of the novel pHrodo Green dye dramatically increases as pH decreases from neutral to acidic, making it an ideal tool to study phagocytosis and its regulation by drugs and/or environmental factors. The lack of fluorescence outside the cell eliminates the need for wash steps and quencher dyes.

Use the ready-made pHrodo Green E. coli BioParticles Conjugate in imaging, high content screening, high throughput screening, and flow applications. Enough reagent is provided to perform 100 tests when using 100 μL in each well of a 96-well plate. The pHrodo Green dye is also available as a conjugate of zymosan BioParticles, S. aureus BioParticles, or dextran 10,000 MW. To create other conjugates, such as antibody conjugates, use pHrodo Green STP ester or pHrodo Green maleimide.

See User Manual for solubility instructions.

Order Info

Shipping Condition: Dry Ice

Specifications

• Cell Type: Mammalian
• Color: Green
• Content And Storage: Store at -20°C, desiccate, and protect from light.
• Description: pHrodo™ Green E. coli BioParticles™ Conjugate for Phagocytosis
• Detection Method: Fluorescence
• For Use With (Application): Cell Analysis
• For Use With (Equipment): Attune Acoustic Focusing Cytometer, Confocal Microscope, Floid Cell Imaging System, Fluorescence Microscope, High Content Analysis Instrument
• Product Type: Conjugate
• Dye Type: pHrodo™ Green
• Form: Solid
• Product Line: pHrodo
• Quantity: 5 x 2 mg
• Shipping Condition: Room Temperature
• Species: E. coli
• Excitation/Emission: 509/533

Frequently Asked Questions (FAQs)

I am performing a phagocytosis assay of macrophages engulfing pHrodo-labeled bacteria. What do you recommend for fixation after the phagocytosis?

pHrodo is relatively non-fluorescent until it enters the acidic phagosome, at which point its fluorescence increases. If you fix the sample, the pHrodo will only reflect the pH of the buffer the cells are in, and not the pH of the phagosome. For this reason, we do not recommend fixing samples. If you want to see how many cells engulfed the labeled bacteria, fix the cells and then place the fixed cells in an acidic buffer for the assay.

Can I store reconstituted pHrodo BioParticles Conjugates for Phagocytosis and Phagocytosis Kit, for Flow Cytometry?

Yes. Once reconstituted, pHrodo BioParticles Conjugates for Phagocytosis and Phagocytosis Kit, for Flow Cytometry (Cat. Nos. P35367, P35361, P35360, P35366, P35364, P35365, A10010) 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. DO NOT FREEZE the resuspended pHrodo bioparticle conjugates.

How many bioparticles are provided in pHrodo Green E. coli BioParticles Conjugate for Phagocytosis (Cat. No. P35366)?

pHrodo Green E. coli BioParticles Conjugate for Phagocytosis (Cat. No. P35366) contains ~3 x 10⁸ bioparticles/mg bioparticles before conjugation. We do not count the number of bioparticles after conjugation.

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.

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.

What can the BioParticles product line be used for?

Fluorescent Bioparticles have been employed to study phagocytosis by fluorescence microscopy, quantitative spectrofluorometry, and flow cytometry. We offer E.Coli, S. aureus, and zymosan BioParticles conjugates covalently labeled with a variety of different fluorophores.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.