Flow cytometry analysis was performed to verify the identification of EVs using Tetraspanin Exo-Flow Combo Capture kit (System Bioscience, cat. no. ExoFlow150A-1) following the manufacturer’s instructions, and standard flow cytometry method (FACS ARIA II, BD). Briefly, tetraspanin (CD9, CD63 and CD81)-biotin antibodies were coupled to the magnetic streptavidin beads of 9.1 μm size. Tetraspanin-coupled magnetic beads were then incubated with isolated EVs (10 μg) to capture EVs, unbound EVs were washed away. Captured EVs were then stained with detection antibodies for CD9 or CD63 conjugated with FITC (System Bioscience, cat. no. ExoFlow 150A-1).
Tetraspanin exo flow combo capture kit
Manufactured by System Biosciences
Sourced in United States
The Tetraspanin Exo-Flow Combo Capture kit is a product designed for the isolation and enrichment of extracellular vesicles (EVs) from biological samples. The kit utilizes magnetic beads coated with antibodies specific to tetraspanin proteins, which are commonly expressed on the surface of EVs. This allows for the selective capture and isolation of EVs from complex samples such as cell culture media or biological fluids.
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Lab products found in correlation
2 protocols using tetraspanin exo flow combo capture kit
1
Tetraspanin-Based EV Identification
2
Detecting P2X7r Expression on Extracellular Vesicles
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Magnetic streptavidin beads were conjugated with tetraspanin-coupled, biotinylated anti-CD9 or anti-CD63 antibodies provided in the Tetraspanin Exo-Flow Combo Capture Kit (System Biosciences, Palo Alto, USA). These magnetic beads were incubated with EV suspension overnight on a rotating mixer at 4 °C. During this step, EVs were captured on to the conjugated magnetic beads. The next morning, magnetic beads were washed thrice in 1X wash buffer to remove any unbound EV particles. EVs captured on to magnetic beads were resuspended in 500 µL wash buffer and incubated with 5 µg of anti-P2X7r antibody conjugated with Alexa Fluor™ 647 overnight on a rotating mixer at 4 °C. The magnetic beads were washed thoroughly to eliminate unbound P2X7r antibodies. EVs were stained with exo-FITC dye (System Biosciences). Cytometric acquisition was performed using an Aurora flow cytometer (Cytek®, San Diego, USA) and analyzed using FlowJo software v10 (Tree Star Inc., Ashland, USA) to check the distribution of P2X7r on EVs.
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