ExoQuick-TC Tissue Culture Media Exosome Precipitation Solution
Exosomes are 60-120 nm membrane vesicles secreted by most cell types in vivo and in vitro. Exosomes are found in blood, urine, amniotic fluid, malignant ascites fluids, tissue culture media and contain different subsets of microRNA and proteins depending on the type of cell from which they are secreted. SBI ExoQuick-TC Exosome Precipitation Reagent is a special polymeric formulation other than the one Original ExoQuick reagent for serum. ExoQuick-TC has been optimized for exosomes isolation of urine samples and media. This technology produces microRNAs and simple, reliable and quantitative protein biomarker discoveries.
Isolate exosomes with ease
• No time-consuming ultracentrifugation
• Less expensive than expensive DynaBeads
• More effective than any other method
• Use as little as 5 ml of cell culture medium or urine
ExoQuick-TC precipitates urinary exosomes
Ten millilitres of normal human urine were pooled with 2 ml of ExoQuick-TC to precipitate urinary exosomes. The exosome pellet was resuspended with 175 μl of buffer and increasing amounts of exosome suspension were loaded onto a plate prepared for ELISA. The CD9 protein was detected using SBI rabbit anti-CD9 primary antibody and SBI’s HRP conjugated goat anti-rabbit secondary antibody. The size of CD9 proteins in urine was determined using Western blot analysis with the same set of antibodies.
NanoSight Analysis of Urine and Tissue Culture Media Exosomes
- Media from HT1080 cells
HT1080 Human Lung Sarcoma Cell Line was grown in a conditioned medium (without serum) for 72 hours. Ten millilitres of the medium was combined with 2 ml ExoQuick-TC to sediment exosomes overnight. The exosome granule was resuspended in 1 ml PBS, diluted at 1:40 and displayed on the NanoSight LM10 instrument. Analysis shows that 133nm Isolated Exosomes with ExoQuick with a recovery of 1.74 x 10 ^ 9 particles/ml.
- HEK293 Cell Media
The human embryonic kidney (HEK) cell line was grown in a conditioned medium (without serum) for 72 hours. Ten millilitres of the medium was combined with 2 ml ExoQuick-TC to sediment exosomes overnight. The exosome granule was resuspended in 1 ml PBS and visualized on the NanoSight LM10 instrument undiluted. Analysis shows that 137nm Exosomes Isolated by ExoQuick with a recovery of 3.33 x 10 ^ 8 particles/ml.
- Human Urine
A normal human urine sample was used. Five millilitres combined with 2.5 ml of ExoQuick-TC to pelletize the exosomes at night. The exosome pellet was resuspended in 1 ml of PBS, diluted at 1:50 and displayed on the NanoSight LM10 instrument. The Analysis shows that ExoQuick isolated 107 nm exosomes with a recovery of 4.80 x 10 ^ 9 particles/ml.