Extracellular vehicles (EVs) are lipid bound vesicles secreted by cells into the extracellular space. The three main subtypes of EVs are microvesicles (100-350 nm), exosomes (30-150 nm), and apoptotic bodies (500-1000 nm), which are differentiated based upon their biogenesis, release pathways, size, content, and function. Exosomes can be formed by extracellular stimulation, microbial attack, and other stress conditions, and are released through either outward budding of the plasma membrane (microvesicle pathway) or inward budding of the endosomal membrane. Inward budding of endosomal membranes results in the progressive accumulation of intraluminal vesicles (ILVs) within the lumen of multivesicular endosomes (MVEs). MVEs fuse with the plasma membrane to release ILVs that are then called exosomes. The regulation of MVE and the formation and release of exosome are mostly mediated by the endosomal sorting complexes required for transport (ESCRT) machinery, or by the ESCRT-independent pathway.

Nat. Rev. Mol. Cell BiolNat. Rev. Mol. Cell Biol. 19, 213–228 (2018).

Exosomes can be released by almost all kinds of cells under physiological and pathological conditions, and are widely distributed through body fluids. Exosomes play a key role in cell communication and epigenetic regulation by transporting proteins and genetic material. Analysis of exosome content to estimate the functional status of parental cells lays a foundation for exosome-based diagnosis. In addition to in disease diagnosis, exosomes are used in a variety of biomedical fields such as drug delivery, vaccine development, and regenerative medicine.

Exosome Detection by Flow Cytometry

Exosomes are difficult to identify because of their small size, strong heterogeneity, and low refractive index. The use of flow cytometry not only enables high-throughput analysis of exosomes, but also allows quantification or classification of exosomes based on their antigenic expression.

Exosomes are immobilized on the surface of the beads, and the exosomal vesicles are exposed to a fluorescently conjugated antibody against an antigen expressed on the exosome surface. When the sample passes through the laser of the flow cytometer, it emits a fluorescent signal which is detected.

Product Range

  • Exosome Detection Kits

Amerigo Scientific offers kits for isolation/detection of exosomes based on the immunobead assay method, which uses a bead-bound capture antibody and a fluorochrome conjugated detection antibody. The kits provide reproducible results and can be run in parallel to exosome immunophenotyping.

Product Name Kit Content Intended Use (RUO)
ExoStepTM Plasma + Standard Superparamagnetic Capture Beads (CD9 Capture Beads)
Primary detection antibody (CD81 PE)
Assay Buffer 10X
Lyophilized exosomes (1x10^12) from Human Plasma
Intended for the detection of human exosomes from Plasma samples
ExoStepTM Culture + Standard Superparamagnetic Capture Beads (CD63 Capture Beads)
Primary detection antibody (CD9 PE)
Assay Buffer 10X
Lyophilized exosomes (1x10^12) from PC-3 Human prostate cancer
Intended for the detection of human exosomes from Cell Culture samples
ExoStepTM Superparamagnetic Capture Beads (CD63 Capture Beads)
Primary detection antibody (CD9 Biotin) or (CD81 Biotin)
Secondary detection reagent (PE Conjugated)
Assay Buffer 10X
Intended for the detection of human exosomes from Plasma or Urine samples
ExoStep Urine Superparamagnetic Capture Beads (CD63 Capture Beads)
Primary detection antibody (CD9 PE)
Assay Buffer 10X
Intended for the detection of human exosomes from Urine samples
ExoStep Plasma Superparamagnetic Capture Beads (CD63 Capture Beads)
Primary detection antibody (CD9 Biotin)
Secondary detection reagent (PE Conjugated)
Assay Buffer 10X
Intended for the detection of human exosomes from Plasma samples
ExoStep Mouse Superparamagnetic Capture Beads (CD63 Capture Beads)
Primary detection antibody (CD9 Biotin)
Secondary detection reagent (PE Conjugated)
Assay Buffer 10X
Intended for the detection of mouse exosomes from Cell Culture samples

Benefits

  • Specific and ambiguous detection
  • Quantitative analysis, and excellent correlation between fluorescence and the amount of exosomes
  • Direct detection of exosomes in cell culture supernatant and biological fluids without isolation or precipitation
  • Small amount of sample needed
  • Greater sensitivity, wide dynamic range, reproducible
  • Allowing simultaneous immunophenotyping of exosomes capture population

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  • Lyophilized Exosome Standards

Compared with fresh exosomes, lyophilized exosomes can be easily transported and stored at low temperature for a long time, and the complete morphology and biological function of exosomes can be maintained after solubilization. In addition to being used as a control standard for flow cytometry, lyophilized exosomes can also be used as a control standard for a variety of applications such as western blot (WB) and ELISA.

Amerigo Scientific offers the highest pure lyophilized exosome standards from human body fluids (plasma and serum) and different cell culture media. The lyophilized standards have been validated by WB and flow cytometry, for overall protein content and particle number by nanoparticles tracking analysis (NTA).


Benefits

  • Highly pure exosomes, providing better performance than competitors
  • Guaranteed stability owing to an exclusive lyophilization procedure
  • Exhaustive validation by WB, NTA, flow cytometry and functional analysis in vitro
  • Tested in application for medicine regenerative, skin, dermatological and pharma companies

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