pHrodo™ BioParticles™ Conjugates for Phagocytosis and Phagocytosis Kit, for Flow Cytometry
pHrodo™ BioParticles™ Conjugates for Phagocytosis and Phagocytosis Kit, for Flow Cytometry
Invitrogen™

pHrodo™ BioParticles™ Conjugates for Phagocytosis and Phagocytosis Kit, for Flow Cytometry

Achieve faster and more accurate results in phagocytosis assays with pHrodo BioParticles conjugates and kit. pHrodo BioParticles conjugates fluoresce brightly in endosomes and lysosomes, do not require wash steps or quencher dye, can be multiplexed, and are used in flow cytometry, HCA, and HTS.
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Catalog NumberColorProductTypeSpecsSpecies
A10025Promo ImageE. coli
P35360E. coli
P35361E. coli
A10010S. aureus
P35367S. aureus
P35365S. cerevisiae
Catalog number A10025
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1,035.65
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1,380.00
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E. coli
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Price (USD)
1,035.65
Online Exclusive
1,380.00
Save 344.35 (25%)
Each
Add to cart
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Achieve faster staining and more accurate results without the need for wash steps or quencher dye with pH-sensitive pHrodo BioParticles conjugates and phagocytosis kit for flow cytometry. pHrodo BioParticles conjugates for phagocytosis are non-fluorescent outside the cell at neutral pH but fluoresce brightly in acidic pH environments, such as those of endosomes and lysosomes. They can be used in imaging, HCA, and HTA applications. The pHrodo BioParticles phagocytosis kit offers rapid assessment of phagocytic activity in whole blood using flow cytometry.

The fluorescence of pHrodo 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. Outside the cell, the dye is non-fluorescent, which eliminates the need for wash steps and quencher dyes. Once internalized, the dye fluoresces brightly within the acidic environments of endosomes and lysosomes, enabling rapid assay development and certainty of results when investigating phagocytic pathways and their regulation by drugs and/or environmental factors. As such, pHrodo dye conjugates can be used in plate readers, fluorescence microscopy imaging, and flow cytometry applications.

The pHrodo Red S. aureus BioParticles Conjugate for Phagocytosis (Cat. No. A10010) offers fast and accurate assay results. pHrodo Red dye conjugates are non-fluorescent outside the cell but fluoresce bright red in phagosomes. The pHrodo Red E. coli BioParticles Conjugate for Phagocytosis (Cat. No. P35361) detects phagocytosis and endocytosis, offers reduced signal variability and improved timing in sensitive experiments, and can be used in multiplex applications with a wide variety of blue, green, and far-red dyes and reporters such as GFP, Fuo-4, calcein, NucBlue, CellEvent Caspase 3/7 green, Mitosox Green, and Mitotracker Deep Red, among many others. The optimal absorption and fluorescence emission maxima of the pHrodo Red BioParticles Conjugate is approximately 560/585 nm, respectively. However, the fluorophore is readily excited with the 488-nm argon-ion laser installed on most flow cytometers.

The pHrodo Red E. coli BioParticles Phagocytosis Kit (Cat. No. A10025) for flow cytometry is designed for rapid and convenient measurement of phagocytic activity in whole blood samples by flow cytometry. The kit includes all the reagents required for assessing particle ingestion and red blood cell lysis.

pHrodo Deep Red dye is a low-background pH sensor dye that shows no signal in neutral conditions and only fluoresces in acidic environments. pHrodo Deep Red dye enables better discrimination of internalized cargo from outside the cell because it has an approximate pKa of 5 and does not fluoresce until it enters the late endosome and lysosome. pHrodo Deep Red dye can be detected using a Cy5 fluorescent filter set and has been validated for use in a variety of applications, including flow cytometry, fluorescent microscopy, high content analysis (HCA), and high throughput screening (HTS). It can also be multiplexed with a wide variety of blue, green, and red dyes and reporters such as GFP and RFPs, Mitosox Green or Red, CellEvent Caspase 3/7 Green or Red, NucBlue, or TMR, among many others.

pHrodo Deep Red E. coli BioParticles Conjugate for Phagocytosis (Cat. No. P35360) is a no-wash, low background, fluorogenic reagent developed for the study of phagocytosis in a live cell system. This conjugate is non-fluorescent outside the cell but fluoresces dark red in phagosomes. The low pKa of the pHrodo Deep Red E. coli BioParticles Conjugate eliminates non-specific signal from non-internalized cargo and only turns on in late endosomes and lysosomes.

The pHrodo Green dye conjugates, like pHrodo Red and pHrodo Deep Red dye conjugates, offer faster and more accurate results than any other phagocytosis assay. pHrodo Green conjugates are non-fluorescent outside the cell at neutral pH but fluoresce bright green at acidic pH, such as that of phagosomes. Use the ready-made pHrodo Green S. aureus BioParticles Conjugate (Cat. No. P35367) and pHrodo Green Zymosan BioParticles Conjugate (Cat. No. P35365) for Phagocytosis in imaging, HCA, HTS, and flow applications.

These pHrodo conjugate products include enough reagent to perform 100 tests when using 100 μL in each well of a 96-well plate, and they can be multiplexed with a wide variety of blue, red, and far-red dye reporters such as Mitosox Red, CellEvent Caspase 3/7 Red, NucBlue, RFPs, and Mitotracker Deep Red, among many others.

pHrodo Red Zymosan BioParticles Conjugate (Cat. No. P35364) is also available, and pHrodo Green dye is available as a conjugate of E. coli BioParticles (Cat. No. P35366).

For Research Use Only. Not for human or animal therapeutic or diagnostic use.
Specifications
DescriptionpHrodo™ Red E. coli BioParticles™ Phagocytosis Kit for Flow Cytometry
Detection MethodFluorescence
Dye TypepHrodo™ Red
FormatBottle(s)
Quantity100 assay kit
Shipping ConditionRoom Temperature
SpeciesE. coli
ColorRed
Emission488
For Use With (Application)Cell Analysis
For Use With (Equipment)Flow Cytometer
Product LineBioParticles, pHrodo
Product TypePhagocytosis Kit
Unit SizeEach
Contents & Storage
• Lysis buffer, 10 mL
• Buffer B, 200 mL
• Wash buffer, 200 mL
• pHrodo™ Red E. coli BioParticles™, 1 vial lyophilized product

Store in refrigerator (2–8°C) and protect from light.

live-cell-tracking-with-phrodo-dyes
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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.

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

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 concentration of bacterial particles for pHrodo BioParticles Conjugates for Phagocytosis?

The bacterial particles per weight for pHrodo BioParticles Conjugates for Phagocytosis, is as follows:

pHrodo E. Coli BioParticles Conjugate for Phagocytosis (Cat. Nos. P35360, P35361, A10025) contains 3 x 108 particles/mg.

pHrodo S. aureus BioParticles Conjugate for Phagocytosis (Cat. Nos. A10010, P35367) contains 3 x 108 particles/mg.

pHrodo Zymosan BioParticles Conjugate for Phagocytosis (Cat. No. P35365) contains 2 x 107 particles/mg.

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

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.

Fluorescence spectra

Fluorescence spectra

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Lot #Certificate TypeDateCatalog Number(s)
2906461Certificate of AnalysisApr 09, 2025A10025
3110702Certificate of AnalysisMar 31, 2025P35361
3059224Certificate of AnalysisMar 13, 2025P35360
3048917Certificate of AnalysisDec 10, 2024P35365
3050023Certificate of AnalysisNov 18, 2024P35367
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Citations & References (80)

Citations & References
Abstract
Extracellular Matrix Lumican Promotes Bacterial Phagocytosis, and Lum-/- Mice Show Increased Pseudomonas aeruginosa Lung Infection Severity.
Authors:Shao H, Lee S, Gae-Scott S, Nakata C, Chen S, Hamad AR, Chakravarti S,
Journal:J Biol Chem
PubMed ID:22865855
'Phagocytosis is central to bacterial clearance, but the exact mechanism is incompletely understood. Here, we show a novel and critical role for lumican, the connective tissue extracellular matrix small leucine-rich repeat proteoglycan, in CD14-mediated bacterial phagocytosis. In Psuedomonas aeruginosa lung infections, lumican-deficient (Lum(-/-)) mice failed to clear the bacterium from ... More
SLAM is a microbial sensor that regulates bacterial phagosome functions in macrophages.
Authors:Berger SB, Romero X, Ma C, Wang G, Faubion WA, Liao G, Compeer E, Keszei M, Rameh L, Wang N, Boes M, Regueiro JR, Reinecker HC, Terhorst C,
Journal:Nat Immunol
PubMed ID:20818396
'Phagocytosis is a pivotal process by which macrophages eliminate microorganisms after recognition by pathogen sensors. Here we unexpectedly found that the self ligand and cell surface receptor SLAM functioned not only as a costimulatory molecule but also as a microbial sensor that controlled the killing of gram-negative bacteria by macrophages. ... More
Innate immunity and transcription of MGAT-III and Toll-like receptors in Alzheimer's disease patients are improved by bisdemethoxycurcumin.
Authors:Fiala M, Liu PT, Espinosa-Jeffrey A, Rosenthal MJ, Bernard G, Ringman JM, Sayre J, Zhang L, Zaghi J, Dejbakhsh S, Chiang B, Hui J, Mahanian M, Baghaee A, Hong P, Cashman J
Journal:Proc Natl Acad Sci U S A
PubMed ID:17652175
'We have tested a hypothesis that the natural product curcuminoids, which has epidemiologic and experimental rationale for use in AD, may improve the innate immune system and increase amyloid-beta (Abeta) clearance from the brain of patients with sporadic Alzheimer''s disease (AD). Macrophages of a majority of AD patients do not ... More
Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming.
Authors:Shin JW, Suzuki T, Ninomiya N, Kishima M, Hasegawa Y, Kubosaki A, Yabukami H, Hayashizaki Y, Suzuki H,
Journal:Nucleic Acids Res
PubMed ID:22879381
'Combinatorial interactions of transcription modulators are critical to regulate cell-specific expression and to drive direct cell reprogramming (e.g. trans-differentiation). However, the identification of key transcription modulators from myriad of candidate genes is laborious and time consuming. To rapidly identify key regulatory factors involved in direct cell reprogramming, we established a ... More
A Review of Reagents for Fluorescence Microscopy of Cellular Compartments and Structures, Part I: BacMam Labeling and Reagents for Vesicular Structures.
Authors:Dolman NJ, Kilgore JA, Davidson MW,
Journal:
PubMed ID:23835803
'Fluorescent labeling of vesicular structures in cultured cells, particularly for live cells, can be challenging for a number of reasons. The first challenge is to identify a reagent that will be specific enough where some structures have a number of potential reagents and others very few options. The emergence of ... More
80 total citations

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