CD63 is frequently used as an exosomal marker.13 (link) We used flow cytometry to study urinary exosomal CD63. We added 1 µL of Aldehyde/sulphate latex beads (A37304, ThermoFisher) to 100 µL of exosome suspension, and mixed well. Nine hundred microliters of PBS was then added, and the mixture was left to incubate for 2 hours at room temperature on a rotary wheel. One hundred microliters of 100 mM glycine was then added and the mixture incubated at room temperature for 30 minutes to block binding. The beads were then pelleted by centrifugation at 2650 g for 5 minutes, and the supernatant removed. The pellet was washed by addition of 1mL PBS and centrifugation at 2650 g for 5 minutes. Finally, the pellet was resuspended in PBS and the antibody of anti-human CD63-FITC FITC (IM1165U, Beckman Coulter) or the antibody of isotype control (A07795, Beckman Coulter) was added according to standard flow cytometry protocol. The antibody was incubated for 30 minute at dark. After twice washing with PBS, the staining was observed by using a conventional flow cytometry. Data were analyzed by using the NaviosTM software (Beckman-Coulter, CA).
Im1165u
Manufactured by Beckman Coulter
The IM1165U is a laboratory instrument manufactured by Beckman Coulter. It is designed to perform automated immunoassay testing. The core function of this product is to detect and quantify specific analytes in biological samples.
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Lab products found in correlation
2 protocols using im1165u
1
Urinary Exosomal CD63 Quantification
2
Exosomal CD63 Expression Analysis
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FCM was utilized to check the exosomal marker CD63. We added 1 µl of Aldehyde/sulphate latex beads (A37304, ThermoFisher) to 200 µl of exosome suspension. The solution was well mixed. Then we added 800 µl of PBS into the solution, and the mixture was left to incubate for 2 h at room temperature on a rotary wheel. The beads were collected by centrifugation at 4000 g for 5 min, and the supernatant was discarded. The beads were washed by the addition of 1 ml PBS and then were centrifuged at 4000 g for 5 min. Finally, the beads were suspended in PBS. The antibody of anti-human CD63-FITC (IM1165U, Beckman Coulter) or the antibody of an isotype control (A07795, Beckman Coulter) was added according to the standard protocol for FCM. The antibody was incubated for 30 min at dark. After twice washing with PBS, the staining was analyzed by using a conventional FCM.
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