ExoStepTM KIT: The most accurate method of exosome detection developed to date.
ExoStepTM kit is the most accurate method of exosome isolation developed to date, a superior alternative for the sensitive detection of exosomes compared with the most commonly used methods. Our kit is easy to implement and analyse for any laboratory that has access to a conventional flow cytometer.
Specific Exosome Detection in biological fluids by Flow Cytometry
Wide dynamic range and limit of detection
Allowing simultaneous Immunophenotyping of captured exosomes population
Easy to perform Immunobead assay with quantitative and scalable results
Specific and unambiguous exosome detection in biological fluids by flow cytometry:
ExoStepTM kit is an Immunobead assay for the detection of exosomes using a bead-bound capture antibody and fluorochrome conjugated detection antibody. This kit provides reproducible results and is intended for Flow Cytometry analysis of pre-enriched human exosomes from biofluids (plasma, serum, urine, among others) or cell culture media.
Ultracentrifugation is not needed
Reproducible results
Effective with small sample quantities
Bead-based assay for Flow Cytometry
ExoStepTM Kit for the Detection and Characterization of exosomes by Flow Cytometry is composed of an exosome capture reagent consisting of 6 µm diameter magnetic beads coated with a specific antibody as solid suport detectable by the cytometer. Additionally, the capture beads are dyed red and fluorescently labelled which facilitate their detection in the fluorescent detector of a conventional flow cytometer. ExoStepTM contains a detector antibody fluorescent conjugated for exosome detection.
Wide dynamic range and limit of detection
To determine the dynamic range and limit of detection of the assay, EVs titration experiments were performed using CD63 for capture and CD9 for detection. The sensitivity of the assay has demonstrated to be very high with a positive signal detected as little as thirty ng of exosomes while 2 ug were required for Western Blot detection. Besides, Flow Cytometry data follows a linear distribution between 30 and 8 micrograms of exosomes, allowing interpolating fluorescent intensity values to estimate the amount of exosomes in the sample.
Simultaneous Immunophenotyping
Considering exosome direct detection results in body fluids, we want to anticipate researcher’s needs for exosome subpopulations characterization through specific markers. In order to optimize the protocol, we performed an Immunophenotyping assay simultaneously with the exosome detection. This optimization of the protocol made possible to include in the FL2 channel (the most sensitive), the antibody against the marker of interest. The suitability of the kit for the specific detection of exosomes was confirmed, making possible to further characterize their phenotype by labelling with other markers of interest.
ExoELISA-Step is an enzyme immunoassay for the detection and quantification of exosomes, based on the use of antigens or antibodies labeled with an enzyme, so that the resulting conjugates have both immunological and enzymatic activity. As one of the components (antigen or antibody) is labeled with an enzyme and insolubilized on a support (immunosorbent), the antigen-antibody reaction will remain immobilized and therefore, is easily revealed by the addition of a specific substrate. When acting on the enzyme a simple or quantifiable view will produce an observable color using a spectrophotometer or a colorimeter.
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