For transmission electron microscopy (TEM), the isolated exosomes were fixed with 1% glutaraldehyde, then a drop of fixed exosomes was spotted onto a formvar/carbon-coated grid and negatively stained with 3% aqueous phosphotungstic acid for 1 min. Subsequently, MSCs-derived exosomes were observed under TEM (Hitachi, Tokyo, Japan, SU-8010). For flow cytometry analysis of exosomes, MSCs-derived exosomes were mixed with 3 μm aldehyde/sulfate latex beads (Invitrogen, Batch Num: 979383) for 10 min with continuous rotation. 1 M glycine in PBS containing 2% BSA was added into the mixture to stop the reaction. Beads coated with exosomes were incubated with antibodies CD63-FITC (Lot: GR320523-9, Abcam), CD81-PE (Cat: MA5-17941, Invitrogen) at 37 ℃ for 25 min. Then a flow cytometry (FCM) (BD FACSCalibur) was used to detect the mesenchymal markers. In addition, the detection of exosomal markers was introduced in western blot assay by using corresponding antibodies including CD9, CD63, CD81, HSP70 and Calnexin.
Cd81 pe
Manufactured by Thermo Fisher Scientific
CD81-PE is a laboratory reagent used for the detection and analysis of CD81 protein expression on the surface of cells. It is a fluorescently labeled antibody that binds to the CD81 antigen, allowing for the identification and quantification of cells expressing this protein using flow cytometry or other cell analysis techniques.
Automatically generated - may contain errors
2 protocols using cd81 pe
1
Characterization of Mesenchymal Stem Cell Exosomes
2
Exosome Characterization via TEM and Flow Cytometry
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For transmission electron microscopy (TEM), isolated exosomes were xed with 1% glutaraldehyde, then a drop of xed exosomes was spotted onto a formvar/carbon-coated grid and negatively stained with 3% aqueous phosphotungstic acid for 1 min. Subsequently, MSCs-derived exosomes were observed under TEM (Hitachi, Tokyo, Japan, SU-8010). For ow cytometry analysis of exosomes, MSCs-derived exosomes were mixed with 3 μm aldehyde/sulfate latex beads (Invitrogen, Batch Num: 979383) for 10 min with continuous rotation. 1M glycine in PBS containing 2% BSA was added into the mixture to stop the reaction. Beads coated with exosomes were incubated with antibodies CD63-FITC (Lot: GR320523-9, Abcam), CD81-PE (Cat: MA5-17941, Invitrogen) at 37 ℃ for 25 min. Then a FCM ow cytometry (BD FACSalibur) was used to detect the mesenchymal markers.
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