MILLIPLEX^® Human Exosome Characterization Panel

Cells release extracellular vesicles (EV), including exosomes, as part of normal physiological processes including removal of waste and regulation of intercellular communication. Furthermore, increased exosome release has been implicated in the progression of various disease states and other abnormalities such as cancer and neurodegeneration. Exosome identification, classification, and differentiation from other extracellular vesicles is often complicated due to the variability in isolation and preparation protocols, exosome biogenesis, as well as exosome source. Thus, development of additional exosome characterization tools is beneficial to the exosome research field. Commonly associated exosome-enriched markers include tetraspanins CD9, CD63, and CD81, cargo sorting-associated markers TSG101 and Flotillin-1, and biogenesis marker syntenin-1. Other markers, including Ago2, GAPDH, and Calreticulin, may or may not be found in exosome-enriched samples, but this can vary depending on source, isolation method, and/or EV biogenesis mechanism. The Human Exosome Characterization panel provides a novel tool for researchers to study exosomal makers and content in lysed exosome samples.

Cross Reactivity: There was no or negligible cross-reactivity between the antibodies for an analyte and any of the other analytes in this panel, except Syntenin-1 and Flotillin-1 antibodies had minimal cross reactivity with GAPDH standard.

This is an overnight assay.

• Each assay requires 25 µL per well of isolated exosomes.
• Argonaute-2 (Ago2) is a qualitative assay with the MCF7:IGF-1 Cell Lysate included as a control. All other analytes are for quantitative analysis.
• To test purity of EV preparation, the MILLIPLEX^® Human Apolipoprotein 6-Plex Panel (Cat. No. APOMAG-62K) can be used.
• To asses the functional component of EVs, the MILLIPLEX^® Human Cytokine/Chemokine Growth Factor Panel A (Cat. No. HCYTA-60K), Panel B (HCYTB-60K), and Multi-Species TGFβ Panel (Cat. No. TGFBMAG-64K-03) can be used.
• Following sample collection and preparation, exosomes can now be isolated and lysed according to your lab’s established protocol. We recommend Cell Signaling Lysis Buffer (Cat. No. 43-040).
• Each lab may use its preferred lysis buffer but must predetermine that the assay will not be negatively affected by the lysis buffer.

Recommended storage for kit components is 2 - 8°C.

Sensitivity: Refer to kit protocol for sensitivities for individual analytes.

MILLIPLEX is a registered trademark of Merck KGaA, Darmstadt, Germany

xMAP is a registered trademark of Luminex Corp

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.