CD9/CD63 Exosome ELISA Kit

The CD9/CD63 Exosome ELISA Kit is a sandwich ELISA kit that utilizes high-performance anti-CD9 (clone 12A12) capture and anti-CD63 (clone 8A12) detection antibodies. It is designed for use on human body fluids or cell culture supernatants to sensitively and quantitatively detect targets that co-express exosome surface markers CD9 and CD63.

Features

• ●Capture exosomes with solid phase anti-CD9 antibody (12A12), then detect using HRP-conjugated anti-CD63 secondary antibody
• ●Directly quantitate exosomes in human blood samples or cell culture supernatants
• ●No special equipment required, apart from a 450nm microplate reader
• ●To implement stability and reproducibility, kit utilizes a CD9/CD63 fusion protein standard instead of unstable/hard-to-store exosomes themselves
• ●Normalization with CD9/CD63 fusion protein standard enables relative quantitation across a set of samples

Data

■Example of measurements for cell culture supernatant

The culture medium containing 10% fatal bovine serum (FBS) was used to culture various cell lines (HCT116, HT29, AsPC-1, MDAMB-231, and PC3). After 8 days of cultivation, supernatants were collected, centrifuged, and then used as samples. Samples were diluted appropriately so that the measurements will be inside the Standard curve window. With this kit, relative quantifi cation of exosomes in the sample will be performed using Standard Protein as a reference.

Based on the measurements for the Standard Protein, plot the absorbance on the X-axis, and the measurements of Standard Protein on the Y-axis (Fig.A). Based on this Standard curve and the sample absorbance, exosome concentration in the samples will be calculated as equivalent to the Standard Protein amount (Fig.B). Since Standard curve will be created for each experiment, one could compare exosome amount directly between the experiments.

■Overview of standardization using CD9/CD63 fusion protein measurement and relative quantification.

The Standard curve was created using the measurement of Standard Protein (CD9/CD63 fusion protein) (Fig.A). From here (Fig.B), assuming 5 pg of CD9/CD63 fusion protein as 1 unit, the OD450 for 1unit will be 0.7. For the exosomes purifi ed from MDM-MB-231, the protein concentration for 1 unit of OD450, which will correspond to 0.7, is about 50ng (Fig.C). Therefore, 50 ng of MDM-MB-231 derived exosomes can be considered as 1unit of CD9 and CD63 positive exosomes. Presenting the exosome measurement by the units, we could standardize it and/or normalize the measurements, and then will be able to compare exosome measurements directly between diff erent samples or diff erent experiments.

Product List

CD9/CD63 Exosome ELISA Kit

Product name	Catalog Number
CD9/CD63 Exosome ELISA Kit, Human	CSR-EXH0102EL