pHrodo™ Red and Green Dextran, 10,000 MW, for Endocytosis
pHrodo™ Red and Green Dextran, 10,000 MW, for Endocytosis
Invitrogen™

pHrodo™ Red and Green Dextran, 10,000 MW, for Endocytosis

Perform fast and accurate live-cell endocytosis and imaging assays with pHrodo Red and Green dextran conjugates, which offer pH-sensitive pHrodo dyes that have no need for washing steps or quencher dye, and which can be multiplexed in imaging, HTS, HCA, tracing, and flow applications.
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Número de catálogoColor
P35368
P10361
Número de catálogo P35368
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Perform live-cell imaging of endocytosis faster and more accurately without the need for wash steps or quencher dye with pHrodo Red and Green dextran 10,000 MW conjugates. The pH-sensitive pHrodo Red and Green dextran conjugates are nonfluorescent in neutral environments but elicit bright red or green fluorescent signal in pH ranges 5–8, such as those in endosomes and lysosomes. These pHrodo-labeled dextrans can be multiplexed with other dyes and used in imaging, HTS, tracing, and flow applications.
pHrodo Red Dextran (Cat. No. P10361) is a superior alternative to other fluorescent dextran conjugates (e.g., BCECF and tetramethylrhodamine [TRITC]) for live-cell imaging of endocytosis, the process whereby the plasma membrane buds to form membrane-bound vesicles (endosomes), which are then trafficked to various destinations within the cell. pHrodo Red Dextran has pH-sensitive fluorescence excitation/emission spectra of 560/585 nm, and its fluorescence emission increases in intensity with increasing acidity. This increase is particularly dramatic in the range pH 5–8, as commonly seen within endocytic vesicles. pHrodo Red Dextran is essentially dark in the extracellular environment; however, upon internalization, the dextran conjugate elicits a bright, red-fluorescent signal in the acidic environment of the endosomes. pHrodo Red Dextran can also be multiplexed 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.

pHrodo Green Dextran (Cat. No. P35368), like pHrodo Red Dextran, offers fast and accurate results for live-cell endocytosis and phagocytosis assays. pHrodo Green conjugates are non-fluorescent outside the cell at neutral pH, but fluoresce bright green at acidic pH ranges, such as those in endosomes and lysosomes. The pHrodo Green dye 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 Green Dextran 10,000 MW conjugate can be used in cell imaging, high content screening, high throughput screening, and flow applications. For a broader selection of compatible dyes, please refer to the Cell Staining Tool or SpectraViewer.

Key applications using labeled dextrans:
There are many citations describing the use of labeled dextrans. Some of the most common uses include the following:
• Neuronal tracing (anterograde and retrograde) in live cells
• Cell lineage tracing in live cells
• Neuroanatomical tracing
• Examining intercellular communications (e.g., in gap junctions, during wound healing, and during embryonic development)
• Investigating vascular permeability and blood-brain barrier integrity
• Tracking endocytosis
• Monitoring acidification (some dextran-dye conjugates are pH-sensitive)
• Studying the hydrodynamic properties of the cytoplasmic matrix.

High manufacturing standards
We offer more than 50 fluorescent and biotinylated dextran conjugates in several molecular weight ranges. Dextrans are hydrophilic polysaccharides characterized by their moderate-to-high molecular weight, good water solubility, and low toxicity. They also generally exhibit low immunogeniticy. Dextrans are biologically inert due to their uncommon poly-(α-D-1,6-glucose) linkages, which render them resistant to cleavage by most endogenous cellular glycosidases.

In most cases, our fluorescent dextrans are much brighter and have higher negative charge than dextrans available from other sources. Furthermore, we use rigorous methods for removing as much unconjugated dye as practical, and then assay our dextran conjugates by thin-layer chromatography to help ensure the absence of low molecular weight contaminants.

Wide selection of substituents and molecular weights
Our dextrans are conjugated to biotin or a wide variety of fluorophores, including seven of our Alexa Fluor dyes and are available in these nominal molecular weights (MW): 3,000; 10,000; 40,000; 70,000; 500,000; and 2,000,000 daltons.

Para uso exclusivo en investigación. No apto para uso diagnóstico o terapéutico en humanos ni en animales.
Especificaciones
DescripciónpHrodo™ Green Dextran, 10,000 MW, for Endocytosis
Método de detecciónFluorescente
Tipo de colorantepHrodo™ Green
Excitación/emisión509/533
FormularioSólido
Cantidad0.5 mg
Condiciones de envíoTemperatura ambiente
EspecieHumano
Localización subcelularEndosomas
ObjetivomiARN
ColorVerde
Para utilizar con (aplicación)Cell Analysis
Para utilizar con (equipo)Citómetro de enfoque acústico Attune™, microscopio confocal, sistema de imágenes celulares Floid™, microscopio de fluorescencia, instrumento de alto contenido, citómetro de flujo, Confocal Microscope, Floid Cell Imaging System, Fluorescence Microscope, High Content Analysis Instrument
Línea de productospHrodo
Tipo de productoSuplemento
Unit SizeEach
Contenido y almacenamiento
Conservar a - 20 °C, desecado y protegido de la luz.

Preguntas frecuentes

I labeled my cells with pHrodo Red dextran dye to examine endocytosis of my adherent cells. After labeling with the recommended concentration in PBS, the cells rounded up and dissociated. Is this a problem with this reagent?

pHrodo Red dextran dye is not known to be toxic to cells. What is more likely is that the cells are not healthy in PBS in the amount of time needed for the assay. Instead, we recommend labeling in a more physiological buffer, such as HBSS, or in phenol red-free media, at an optimal temperature for your cells.

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

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.

What is the optimal working concentration for pHrodo Red and Green Dextran, 10,000 MW, for Endocytosis (Cat. Nos. P10361, P35368)?

The working concentration for pHrodo Red and Green Dextran, 10,000 MW, for Endocytosis (Cat. Nos. P10361, P35368) is anywhere between 10 and 100 µg/mL, depending on cell type, level of background, duration of loading, and the detection modality. We recommend starting with 50 µg/mL treatment and then titrating up or down to get an optimal signal.

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

Citations & References (21)

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
Loss of PiT-1 results in abnormal endocytosis in the yolk sac visceral endoderm.
Authors:Wallingford MC, Giachelli CM
Journal:
PubMed ID:25138534
'PiT-1 protein is a transmembrane sodium-dependent phosphate (Pi) transporter. PiT-1 knock out (KO) embryos die from largely unknown causes by embryonic day (E) 12.5. We tested the hypothesis that PiT-1 is required for endocytosis in the embryonic yolk sac (YS) visceral endoderm (VE). Here we present data supporting that PiT-1 ... More
Walking nanothermometers: spatiotemporal temperature measurement of transported acidic organelles in single living cells.
Authors:Oyama K, Takabayashi M, Takei Y, Arai S, Takeoka S, Ishiwata S, Suzuki M
Journal:Lab Chip
PubMed ID:22437040
We fabricated fluorescent nanoparticles which monitor temperature changes without sensitivity to pH (4-10) and ionic strength (0-500 mM). The nanothermometers spontaneously enter living HeLa cells via endocytosis, enclosed in acidic organelles, i.e., endosome/lysosome, and then transported along microtubules in a temperature-dependent manner, working as "walking nanothermometers". ... More
A New Role for Myosin II in Vesicle Fission.
Authors:Flores JA, Balseiro-Gomez S, Cabeza JM, Acosta J, Ramirez-Ponce P, Ales E,
Journal:
PubMed ID:24959909
An endocytic vesicle is formed from a flat plasma membrane patch by a sequential process of invagination, bud formation and fission. The scission step requires the formation of a tubular membrane neck (the fission pore) that connects the endocytic vesicle with the plasma membrane. Progress in vesicle fission can be ... 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