ExoQuick precipitation kit (System Biosciences, Mountain View, CA, USA) was used to extract exosomes from cell culture medium or serum samples. In brief, Cells were collected when reached 80% confluency and centrifuged at 3000×g for 10 minutes to eliminate cell debris. Then, 63 μL ExoQuick precipitation kit was mixed with 250 μL supernatant and chilled at 4°C for 30 min. The Mixtures were centrifuged at 1500×g for 30 min. After the supernatant was removed, the pellet was re-centrifuged at 1500×g for 5 min to eliminate the residual liquid. The exosome was re-suspended with 200 μL phosphate-buffered saline (PBS). All centrifugations were performed at 4°C.
Exoquick precipitation kit
Manufactured by System Biosciences
Sourced in United States
The ExoQuick precipitation kit is a product designed for the isolation and purification of extracellular vesicles, including exosomes, from various biological samples such as cell culture media or body fluids. The kit utilizes a proprietary polymer-based precipitation method to effectively concentrate and recover these vesicles from the sample.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Beyotime (6 mentions)
Pkh26 red fluorescent cell linker kit for general cell membrane labeling, by Merck Group (6 mentions)
Phosphate buffered saline (pbs), by Solarbio (6 mentions)
Ht7700, by Hitachi (3 mentions)
2100 bioanalyzer, by Agilent Technologies (3 mentions)
Amicon ultra centrifugal filter unit, by Merck Group (3 mentions)
Phosphate buffered saline pbs, by Beyotime (3 mentions)
Ab2787, by Abcam (3 mentions)
Ab30871, by Abcam (3 mentions)
Jem 1010, by JEOL (3 mentions)
Jem 1400, by JEOL (3 mentions)
Zetasizer, by Malvern Panalytical (3 mentions)
Copper grid, by Agar Scientific (3 mentions)
Gene pulser x electroporator, by Bio-Rad (1 mentions)
33 protocols using exoquick precipitation kit
1
Exosome Extraction from Cell Culture
2
Serum Small Extracellular Vesicle Isolation
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Exosome-enriched serum sEV fractions were precipitated using ultracentrifugation and an ExoQuick precipitation kit (System Biosciences Inc., Mountain View, CA, United States)[16 (link)]. The sizes and particle concentrations of the isolated serum sEVs were measured by nanoparticle tracking analysis (NTA, NanoSight NS300, Malvern, United Kingdom). Serum sEVs were visualized using transmission electron microscopy (TEM, HT7700, Hitachi Ltd., Tokyo, Japan). The expression of exosomal protein markers was determined by Western blot analysis. The details are provided in the Supplementary material , Supporting Information.
3
Exosome Extraction from Cell Culture
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Exosome extraction from cell culture medium was performed with ExoQuick precipitation kit (System Biosciences, USA). Briefly, cells were collected when 80% confluency was achieved. After 10 minutes’ centrifugation at 3000 g, cells and cell debris were discarded. Thereafter, 250 μL supernatant was mixed with 63 μL ExoQuick precipitation kit and then incubated for 30 minutes at 4°C. After centrifugation at 1500 g for 30 minutes, the supernatant was discarded. Subsequently, the residual liquid was removed by centrifugation at 1500 g for 5 minutes to obtain the final exosome pellets.
4
Exosome Isolation from Serum Samples
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Exosomes were extracted from serum samples using an ExoQuick precipitation kit (SBI; System Biosciences, Mountain View, CA, USA) according to the manufacturer’s instructions. In brief, serum was thawed on ice and centrifuged at 3,000 × g for 15 min to remove cells and cell debris. Next, 250 μL of the supernatant was mixed with 63 μL of the ExoQuick precipitation kit and incubated at 4°C for 30 min, followed by centrifugation at 1,500 × g for 30 min. Then, the supernatant was removed by careful aspiration, followed by another 5 min of centrifugation to remove the residual liquid. The exosome-containing pellet was subsequently re-suspended in 250 μL PBS. Exosomes were measured for their protein content using a bicinchoninic acid (BCA) protein assay kit (Pierce, Rockford, USA). Electron microscopy was applied to characterize the vesicles floated in PBS.
5
Serum Exosome Isolation and Characterization
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The serum-exosomes were isolated with ExoQuick precipitation kit (EXOQ5A-1; System Biosciences) according to the manufacturer's instructions. Briefly, ExoQuick solution was added into the serum samples and incubated for 30 min at 4°C. Then the mix was centrifuged for 30 min at 1,500 × g at room temperature. Subsequently, the supernatant was carefully removed and then centrifuged for 5 min at 1,500 × g to remove the extra liquid. Exosome pellets were resuspended in PBS (P1022; Beijing Solarbio Science & Technology Co., Ltd.) and preserved at −80°C. Exosomes were isolated from cultured cells through differential ultracentrifugation as previously described (24 (link)).
6
Exosome Isolation from Serum Samples
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Exosomes were extracted from serum samples using an ExoQuick precipitation kit (SBI, System Biosciences, Mountain View, CA), according to the manufacturer’s instructions. Briefly, serum was thawed on ice and centrifuged at 3,000 × g for 15 min to remove cells and cell debris. Next, 250 μL supernatant was mixed with 63 μL ExoQuick precipitation kit and incubated at 4°C for 30 min, followed by centrifugation at 1,500 × g for 30 min. Then, the supernatant was removed by careful aspiration, followed by another 5 min of centrifugation to remove the residual liquid. The exosome-containing pellet was subsequently re-suspended in 250 μL PBS. Exosomes were measured for their protein content using a BCA protein assay kit (Pierce, Rockford, IL, USA). Electron microscopy was applied to characterize the vesicles floated in PBS.
7
Exosome Extraction from Cell Culture and Serum
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ExoQuick precipitation kit (System Biosciences, Mountain view, CA, USA) was used to extract exosomes from GC cells culture medium or serum samples. In brief, the culture medium and serum were unfrozen on ice, followed by centrifugation (3000 × g, 15 min) to discard the cell debris. Then, the supernatant (250 µL) was mixed with the ExoQuick exosome precipitation solution (63µL) and incubated at 4°Cfor 40 min. Next, the supernatant was carefully discarded by aspiration following centrifugation (1500 × g, 15 min), followed by another centrifugation (1500 × g, 5mins) to remove the residual liquid. Subsequently, phosphate buffered saline (PBS, 250 μL) was employed to resuspend the exosome containing pellet. Finally, the pellets containing exosomes were harvested for RNA isolations.
8
Exosomal RNA Extraction from Plasma
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The clinical specimens were thawed on ice. Then, exosomes were isolated from the plasma using an exoQuick precipitation kit (System Biosciences, USA) which was used according to the manufacturer's protocol. The total RNA, including the miRNAs, was extracted using the RNA Isolation Reagent kit (Vazyme, Nanjing, China) and was used according to manufacturer's instructions. In addition, after adding isopropyl alcohol, 1 μg of glycogen was required for each sample. After this step, the mixture was stored at -80°C for overnight to precipitate small RNA. The purity and concentration were determined using Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA).
9
Extracellular Vesicle Isolation from Cell and Plasma
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EVs were extracted from HAEC cell culture medium or plasma samples using an ExoQuick precipitation kit (SBI, System Biosciences, Mountain view, CA) according to the manufacturer’s instructions. Briefly, the culture medium and plasma were thawed on ice and centrifuged at 3000×g for 15 min and 10,000×g for 30 min. For plasma EV isolation, 250 μl of the supernatant was mixed with 67 μl of the ExoQuick precipitation reagent and incubated at 4°Cfor 30 min, followed by centrifugation at 3000×g for 10 min. For the isolation of EVs in cell medium, an Amicon Ultra Centrifugal Filter Unit (100 kDa, Millipore) was used to concentrate the supernatant. The ultrafiltration supernatant was mixed with the ExoQuick precipitation reagent at the ratio of 5:1, and incubated at 4 °C overnight, followed by centrifugation at 1500×g for 30 min. The EV pellet was subsequently resuspended in 200 μl phosphate buffered saline (PBS). This isolation method has been well validated with other techniques including electron microscopy [20 (link)]. EV concentrations and size distribution were measured by nanoparticle tracking analysis (NTA) (NanoSight, NanoSight Ltd., UK).
10
Exosome Isolation from Serum Samples
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Serum exosomes were isolated from serum samples using ExoQuick precipitation kit (Systembiosciences, Mountain View, CA, USA). Firstly, the serum was centrifuged at 3000×g for 15 min to eliminate cell debris. Then, the supernatant was mixed with ExoQuick precipitation kit and centrifuged at 1500×g for 30 min, respectively. Next, the exosomes were centrifuged at 1500×g for 5 min to remove the residual liquid. Finally, the exosomes were re-suspended in phosphate-buffered saline (PBS).
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