Serum exosome levels were determined by a CD9/CD63 exosome ELISA kit (Cosmo Bio., Tokyo, Japan), which is a sandwich ELISA kit that uses anti-CD63 antibody as a capture antibody and anti-CD9 antibody as a detection antibody, according to the manufacturer’s instructions.
Cd9 cd63 exosome elisa kit
Manufactured by Cosmo Bio
Sourced in Japan
The CD9/CD63 exosome ELISA kit is a laboratory assay used to detect and quantify the presence of exosomes expressing the CD9 and CD63 surface markers. The kit provides a standardized method for the specific capture and measurement of these exosome subpopulations from biological samples.
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Lab products found in correlation
3 protocols using cd9 cd63 exosome elisa kit
1
Exosomal Biomarker Quantification by ELISA
2
Isolation and Characterization of Extracellular Vesicles from 293T Cells
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293 T cells were cultured in HE100 medium (Gmep Incorporated, Kurume, Fukuoka, Japan) with L-glutamine. Scramble, miR-709, and miR-1247-3p mimics were synthesized by Ajinomoto Bio-Pharma Services, GeneDesign Inc. (Osaka, Japan). 293 T cells were seeded in a 10-cm dish (2 × 106 (link) cells/dish) by using 10 mL of HE100 medium supplemented with L-glutamine. The next day, the 293 T cells were transfected with 600 pmol of miRNA mimics by using 25 μL of the RNAiMAX transfection reagent (#13778075; Thermo Fisher Scientific), and were incubated for 24 h after transfection. After removing the culture medium from the dishes, 10 mL/dish of fresh HE100 medium supplemented with L-glutamine was applied to the dishes. The culture medium was collected after 3 days. The culture medium was concentrated with Amicon Ultra-15 (#UFC901024, Merck Millipore Ltd.), and the concentrated solution was applied to a qEV2 70 column (Meiwafosis Co., Ltd., Tokyo Japan) in order to isolate the EVs. The EV concentration was measured by using a CD9/CD63 exosome ELISA kit (#EXH0102EL; Cosmo Bio Co., Ltd., Tokyo, Japan).
3
Isolation and Characterization of Tumor-Derived Extracellular Vesicles
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SAS and SAS‐R cells (1 × 106) were seeded in 100 mm tissue culture dishes (AGG Inc., Tokyo, Japan). After 48 h, the conditioned medium (CM) was replaced with 10 ml DMEM containing 10% Exo‐FBS Exosome‐depleted FBS (System Biosciences (SBI), Palo Alto, CA, USA). After a further 24 h, CM was collected and ultrafiltered with a 100‐kDa cut‐off Ultrafiltration filter (Amicon Ultra‐15; MERCK, Tokyo, Japan). The CM (10 ml) was concentrated to approximately 500–700 μl and loaded on a size‐exclusion chromatography (SEC) column (EVSecond L70; GL Sciences, Tokyo, Japan).The fractions were collected according to the manufacturer's protocol. The EV elution fractions were examined by western blotting for CD9, CD81, and ALIX. Moreover, relative quantification of the exosomes was performed using a CD9/CD63 exosome ELISA kit (COSMO BIO, Tokyo, Japan), which could measure exosomes comprising a combination of CD9 and CD63. Furthermore, serum protein concentration was measured using the TaKaRa Bradford Protein Assay Kit (Takara Bio). Subsequently, the eluted samples were pooled. EVs were recovered in phosphate‐buffered saline (PBS) and stored at −80°C. EVs isolated from SAS cells (SAS EVs) and EVs isolated from SAS‐R cells (SAS‐R EVs) were used in subsequent experiments.
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