Exosomes were isolated from media conditioned by human myeloma cell lines. Cells were washed thrice with PBS and grown in serum-free medium for 16 h. Exosomes secreted into the medium by myeloma cells were isolated by differential centrifugation and characterized as described previously [22 (link)]. For exosomes isolated from myeloma patient sera, samples were obtained from a multiple myeloma patient or healthy control enrolled in the Molecular And Genetic Epidemiology (iMAGE) study of myeloma who met the revised and updated International Multiple Myeloma Working Group classification criteria for myeloma [55 (link), 56 (link)]. Approval from the Institutional Review Board in accordance with the Declaration of Helsinki was obtained prior to study initiation, and informed consent was obtained from all individual participants included in the study. Exosomes were isolated from myeloma patient sera using an ExoQuick isolation kit (System Biosciences). Briefly, 30 μl of ExoQuick solution was added to 100 μl of serum and incubated at 4°C for 1 h followed by centrifugation at 1500 × g for 30 min. Particle size and number were assessed using a NanoSight 300 according to manufacturer’s protocol.
Exoquick isolation kit
Manufactured by System Biosciences
Sourced in United States
The ExoQuick isolation kit is a product offered by System Biosciences for the isolation of extracellular vesicles, such as exosomes, from a variety of sample types, including cell culture media and biological fluids. The kit uses a proprietary polymer-based precipitation method to isolate and concentrate extracellular vesicles from the sample.
Automatically generated - may contain errors
4 protocols using exoquick isolation kit
1
Isolation and Characterization of Myeloma Exosomes
2
Exosomal circRNA Profiling in Lung Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasma exosomes were extracted from lung adenocarcinoma samples and PC9/CXCR4-shRNA, PC9/shCtrl, and PC9 cells by using an ExoQuick Isolation kit (System Biosciences, USA).
Total RNA was subsequently extracted from these samples with a QIAGEN RNeasy Mini kit (Qiagen, Duesseldorf, Germany), according to the manufacturer's instructions. Real-time quantitative reverse transcription-polymerase chain reactions (RT-qPCR) were performed with SYBR® Premix Ex TaqTM II (Tli RNaseH Plus, TaKaRa, Dalian, China), according to the manufacturer's instructions. Divergent primers for exo-hsa_circRNA_0056616 and primers for CXCR4 and GAPDH, which was to be the reference gene, were designed by Biocan (Shenzhen, China) and synthesized by TaKaRa. The normalization of qPCR in the expression of exo-hsa circRNA 0056616 was referring to Wang's methods 13 (link). All RT-qPCR primer sets are listed inTable S1 and S2.
Total RNA was subsequently extracted from these samples with a QIAGEN RNeasy Mini kit (Qiagen, Duesseldorf, Germany), according to the manufacturer's instructions. Real-time quantitative reverse transcription-polymerase chain reactions (RT-qPCR) were performed with SYBR® Premix Ex TaqTM II (Tli RNaseH Plus, TaKaRa, Dalian, China), according to the manufacturer's instructions. Divergent primers for exo-hsa_circRNA_0056616 and primers for CXCR4 and GAPDH, which was to be the reference gene, were designed by Biocan (Shenzhen, China) and synthesized by TaKaRa. The normalization of qPCR in the expression of exo-hsa circRNA 0056616 was referring to Wang's methods 13 (link). All RT-qPCR primer sets are listed in
3
Serum Extracellular Vesicle Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
). An ExoQuick isolation kit (System Biosciences) was used in the present study. ExoQuick ULTRA was added to 250 mL serum and the mixture was incubated at 4 °C for 30 min. Then the sample was centrifuged at 3000×g for 10 min. The resulting EVs were resuspended and transferred to a pre-washed ExoQuick ULTRA column. Then the column was centrifuged at 1000×g for 30 s.
4
Extracellular Vesicle Isolation and Characterization
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!