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

Citas y referencias (16)

Invitrogen™

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

Name: pHrodo™ BioParticles™ Conjugates for Phagocytosis and Phagocytosis Kit, for Flow Cytometry
SKU: P35365
Price: 832.68 USD

Obtenga resultados más rápidos y exactos en ensayos de fagocitosis con los conjugados y el kit pHrodo BioParticles. Los conjugados pHrodo BioParticles muestran una fluorescencia brillante en los endosomas y lisosomas, no requieren pasos de lavado ni colorantes supresores, se pueden multiplexar y se utilizan en citometría de flujo, HCA y HTS.

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Número de catálogo	Color	ProductTypeSpecs	Especie
P35365			S. cerevisiae
P35360			E. coli
P35361			E. coli
A10025			E. coli
A10010			S. aureus
P35367			S. aureus

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Número de catálogo P35365

Precio (USD)

832,68

Each

Añadir al carro de la compra

Especie:

S. cerevisiae

Precio (USD)

832,68

Each

Añadir al carro de la compra

Consiga una tinción más rápida y resultados más exactos sin necesidad de pasos de lavado o colorantes supresores con conjugados pHrodo BioParticles sensibles al pH y kit de fagocitosis para citometría de flujo. Los conjugados pHrodo BioParticles para fagocitosis no son fluorescentes fuera de la celda con pH neutro, pero muestran fluorescencia brillante en ambientes con pH ácido, como los de los endosomas y lisosomas. Se pueden utilizar en aplicaciones de adquisición de imágenes, HCA y HTA. El kit de fagocitosis pHrodo BioParticles ofrece una evaluación rápida de la actividad fagocítica en sangre completa mediante citometría de flujo.

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.

Especificaciones

DescripciónConjugado para fagocitosis pHrodo™ Green Zymosan BioParticles™

Método de detecciónFluorescente

Tipo de coloranteColoraciones sensibles al pH

Excitación/emisión509/533

Cantidad5 x 1 mg

Condiciones de envíoTemperatura ambiente

EspecieS. cerevisiae

ColorVerde

Para utilizar con (aplicación)Análisis celular

Para utilizar con (equipo)Citómetro de focalización acústica Attune™, Microscopio confocal, Sistema de adquisición de imágenes de células Floid™, Microscopio de fluorescencia, Instrumentos de alto contenido, Citómetro de flujo

Línea de productospHrodo

Tipo de productoConjugado

Unit SizeEach

Contenido y almacenamiento

Almacenar a – 20 °C, desecar y proteger de la luz.

Preguntas frecuentes

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.

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 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.

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.

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

Citations & References (16)

Citations & References

Abstract

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

A modern descendant of early green algal phagotrophs.

Authors:Maruyama S, Kim E,

Journal:

PubMed ID:23707430

Green algae, land plants, and other photosynthetic eukaryotes possess plastids, such as chloroplasts, which have evolved from cyanobacterial ancestors via endosymbiosis [1]. An early evolutionary merger between heterotrophic eukaryotes and cyanobacteria called primary endosymbiosis gave rise to the first photosynthetic eukaryotes. A series of plastid acquisitions involving engulfment of eukaryotic phototrophs, ... More

Glutathione reductase is essential for host defense against bacterial infection.

Authors:Yan J, Ralston MM, Meng X, Bongiovanni KD, Jones AL, Benndorf R, Nelin LD, Joshua Frazier W, Rogers LK, Smith CV, Liu Y,

Journal:Free Radic Biol Med

PubMed ID:23623936

Glutathione reductase (Gsr) catalyzes the reduction of glutathione disulfide to glutathione, a major cellular antioxidant. We have recently shown that Gsr is essential for host defense against the gram-negative bacteria Escherichia coli in a mouse model of sepsis. Although we have demonstrated that Gsr is required for sustaining the oxidative ... More

NFAT signaling in human mesenchymal stromal cells affects extracellular matrix remodeling and antifungal immune responses.

Authors:

Journal:iScience

PubMed ID:34195564

Deficiency of Microglial Autophagy Increases the Density of Oligodendrocytes and Susceptibility to Severe Forms of Seizures.

Authors:

Journal:Eneuro

PubMed ID:33472865

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