Exoquick ultra ev isolation kit

Manufactured by System Biosciences

Sourced in United States

The ExoQuick ULTRA EV Isolation Kit is a product offered by System Biosciences. It is designed to isolate and purify extracellular vesicles (EVs) from various biological samples, including cell culture media and body fluids. The kit uses a proprietary precipitation-based method to capture and concentrate EVs, which can then be used for downstream applications such as analysis or further processing.

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Lab products found in correlation

Qubit 3.0 fluorometer, by Thermo Fisher Scientific (9 mentions) Ripa buffer, by Thermo Fisher Scientific (6 mentions) Dynabeads m 270 amine, by Thermo Fisher Scientific (3 mentions) Pkh26 fluorescent dye, by Merck Group (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Interleukin 6 (il 6), by Merck Group (1 mentions) Halt protease and phosphatase inhibitor single use cocktail, by Thermo Fisher Scientific (1 mentions) Nonidet p 40, by Merck Group (1 mentions)

11 protocols using exoquick ultra ev isolation kit

Isolation of Serum-Derived Extracellular Vesicles

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EVs were isolated from serum samples using the ExoQuick ULTRA EV isolation kit (System Biosciences, Cat #EQULTRA-20A-1, USA) following manufacturer instructions as previously described (Coughlan et al., 2020 (link)). Isolated EVs were stored at −80°C until analysis.

Laso-García F., Piniella D., Gómez-de Frutos M.C., Casado-Fernández L., Pérez-Mato M., Alonso-López E., Otero-Ortega L., Bravo S.B., Chantada-Vázquez M.D., Trilla-Fuertes L., Fresno-Vara J.Á., Fuentes B., Díez-Tejedor E., Gutiérrez-Fernández M, & Alonso De Leciñana M. (2023). Protein content of blood-derived extracellular vesicles: An approach to the pathophysiology of cerebral hemorrhage. Frontiers in Cellular Neuroscience, 16, 1058546.

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Immunomagnetic Bead-based A673 EV Isolation

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For immunomagnetic bead separation, Tz-grafted magnetic beads were prepared by incubating 2.8 μm Dynabeads M-270 Amine (2 × 10⁸ beads, 100 μL, Thermo Fisher Scientific) with Tz-sulfo-NHS ester (0.32 mg, Click Chemistry Tools, USA) in PBS buffer for 1 h at room temperature. Each artificial A673 EV plasma sample was pre-incubated with the TCO-anti-LINGO-1 conjugate (1 pmol) for 20 min and incubated with Tz-grafted magnetic beads (2 × 10⁷ beads) at room temperature for 30 min to isolate A673 EVs. For ultracentrifugation, each artificial A673 EV plasma sample was centrifuged at 100 000 g for 2 h using Optima L-100 XP Ultracentrifuge. For the commercially used EV isolation assay, each artificial A673 EV plasma sample was isolated and purified using the ExoQuick ULTRA EV Isolation Kit (System Biosciences) according to the manufacturer’s protocol. For all the methods, RNA was extracted from the isolated EVs and quantified using Qubit 3.0 Fluorometer (Thermo Fisher Scientific), followed by quantification of EWS-FLI1 type 1 rearrangement using RT-ddPCR detection. Healthy-donor plasma samples without A673 EVs were processed in parallel to give the systems’ RNA background.

Dong J., Zhang R.Y., Sun N., Hu J., Smalley M.D., Zhou A., Yue H., Rothermich W., Chen M., Chen J., Ye J., Teng P.C., Qi D., Toretsky J.A., Tomlinson J.S., Li M., Weiss P.S., Jonas S.J., Federman N., Wu L., Zhao M., Tseng H.R, & Zhu Y. (2020). Coupling Nanostructured Microchips with Covalent Chemistry Enables Purification of Sarcoma-Derived Extracellular Vesicles for Downstream Functional Studies. Advanced functional materials, 30(49), 2003237.

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Plasma EV Isolation via ExoQuick Kit

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Plasma EV samples used in Fig. 3b were isolated with an ExoQuick ULTRA EV Isolation Kit (EQULTRA-20A-1, System Biosciences) following manufacturer instructions. Briefly, 250 μL plasma aliquots were centrifuged at 3000g for 15 minutes, supernatants were then gently mixed with 67 μL ExoQuick solution and incubated on ice for 30 minutes, and then centrifuged at 3000g and 4 °C, for 10 minutes. EV pellets were then processed according to the manufacturer’s instructions, and EVs was then analyzed by NanoSight.

Ning B., Huang Z., Youngquist B.M., Scott J.W., Niu A., Bojanowski C.M., Zwezdaryk K.J., Saba N.S., Fan J., Yin X.M., Cao J., Lyon C.J., Li C.Z., Roy C.J, & Hu T.Y. (2021). Liposome-mediated detection of SARS-CoV-2 RNA-positive extracellular vesicles in plasma. Nature nanotechnology, 16(9), 1039-1044.

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Exosome Isolation from Mouse Plasma

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Exosomes were extracted from mouse plasma samples and purified with the ExoQuick® ULTRA EV isolation kit, which was purchased from System Biosciences (Mountain View CA, USA). According to the manufacturer’s instruction, 125 μL of plasma was pre-treated with 1 μL of Thrombin to remove fibrin and then incubated with 33.5 μL of ExoQuick exosome precipitation solution for 30 min at 4°C. After centrifugation at 3,000 g for 10 min, the exosomes were concentrated at the bottom of the tubes and then resuspended with buffer. Finally, exosome lysates were prepared by adding RIPA buffer (Thermo Fisher Scientific) and analyzed by Western Blot.

Yi B., Cheng H., Wyczechowska D., Yu Q., Li L., Ochoa A.C., Riker A.I, & Xi Y. (2021). Sulindac modulates the response of proficient MMR colorectal cancer to anti-PD-L1 immunotherapy. Molecular cancer therapeutics, 20(7), 1295-1304.

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Exosome Isolation from Bronchoalveolar Lavage Fluid

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Exosomes isolated from bronchoalveolar lavage fluid (BALF) were centrifuged at 3000× g for 15 min to remove cellular debris, and then subsequently transferred into a new tube. The appropriate amount of ExoQuick solution was added incubated with the BALF using a commercially available kit, ExoQuick ULTRA EV Isolation Kit (System Biosciences, Palo Alto, CA, USA). The solution and the BALF were mixed well and incubated overnight. The ExoQuick/BALF mixture was centrifuged at 1500× g for 30 min. Centrifugation was performed at room temperature or 4 °C. The supernatant was carefully aspirated following centrifugation at 1500× g for 5 min. The remaining, precipitated EV pellet was resuspended in 350 µL of Lysis Buffer, vortexed for 15 s, and allowed to sit at room temperature for 5 min. The quantification and the confirmation of exosomes in the sample were conducted using Nanocyte technology according to previously published literature [34 (link)].

Lopez K., Camacho A., Jacquez Q., Amistadi M.K., Medina S, & Zychowski K. (2022). Lung-Based, Exosome Inhibition Mediates Systemic Impacts Following Particulate Matter Exposure. Toxics, 10(8), 457.

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Plasma EV Isolation via ExoQuick Kit

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Isolation of Glioblastoma-Derived Extracellular Vesicles

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Cells were cultured in 10% EV-FBS-free DMEM (A2720801, GIBCO BRL, Green Island, NY, USA) under normoxia (21% O 2 ) or hypoxia (1% O 2 ). After 48-72 h of incubation, the culture medium was collected and the EVs were separated by ultracentrifugation. The EVs were isolated following the detailed procedures as previously described [7] . Human glioblastoma-EVs or murine glioblastoma-EVs were isolated using the Exoquick ULTRA EV Isolation Kit (EQULTRA-20A-1, System Biosciences, Irvine, CA, USA).

Zhao D., Yu H., Ding L., Wang W., Wang H., Hu Y., Qin L., Deng G., Xie B., Li G., & Qi L. (2020). Glioblastoma cell-derived extracellular vesicle miR-27a-3p facilitates M2 macrophage polarization.

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Exosome Isolation and Characterization from IL-6-Treated hUC-MSCs

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hUC-MSCs were cultured with 1 ng/mL IL-6 (Sigma-Aldrich) in the medium for 48 h, and the supernatant was collected for exosome purification using an ExoQuick ULTRA EV isolation kit (SBI, Palo Alto, CA, USA). The characterization of exosomes was confirmed by electron microscopy and particle size by NanoSight analysis (Shanghai Oe Biotech Co., Ltd.) as described [19 (link)]. The expression of the exosome-specific marker TSG101 (SBI) and the EV-related protein markers CD63 and CD81 (SBI) were analyzed by western blotting. miRNAs were extracted from exosomes using TRIzol reagent (Invitrogen, USA) for qPCR analysis or sequencing at Shanghai Oe Biotech. To identify the transport of exosomes, exosomes were labeled with PKH26 fluorescent dye (Sigma-Aldrich) [19 (link)].

Shao M., Xu Q., Wu Z., Chen Y., Shu Y., Cao X., Chen M., Zhang B., Zhou Y., Yao R., Shi Y, & Bu H. (2020). Exosomes derived from human umbilical cord mesenchymal stem cells ameliorate IL-6-induced acute liver injury through miR-455-3p. Stem Cell Research & Therapy, 11, 37.

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Exosome Isolation from Serum

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The serum samples were thawed at 4°C and then used to isolate exosomes with ExoQuick® ULTRA EV Isolation Kit (SBI, USA) according to the manual's instruction. Briefly, the serum samples were centrifuged at 3000 g for 15 min and then centrifuged at 12000 g for 10 min to remove cellular debris. All the centrifugation was performed at 4°C. Subsequently, 67 μL of ExoQuick was added to 250 μL of the centrifuged serum and incubated at 4°C for 30 min. The pellets of exosomes were collected by centrifuging the ExoQuick/serum mixture at 3000 g for 10 min. After that, the isolated exosomes were purified by elution of purification columns. If the purified exosomes were not immediately used for experiments, they should be stored at -80°C and avoid repeated freezing and thawing.

Zhang P., Liang T., Chen Y., Wang X., Wu T., Xie Z., Luo J., Yu Y, & Yu H. (2020). Circulating Exosomal miRNAs as Novel Biomarkers for Stable Coronary Artery Disease. BioMed Research International, 2020, 3593962.

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Plasma Exosome Isolation and Purification

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Plasma exosomes were isolated and purified with ExoQuick ULTRA EV Isolation Kit (SBI System Biosciences, Palo Alto, CA), according to the manufacturer’s protocol. Briefly, 67 µL of ExoQuick was added to 250 µL plasma after debris was removed and then incubated on ice for 30 min and centrifuged at 3000× g for 10 min. The pellet was resuspended and loaded to a column for purification.

Yao Y., Subedi K., Liu T., Khalasawi N., Pretto-Kernahan C.D., Wotring J.W., Wang J., Yin C., Jiang A., Fu C., Dimitrion P., Li J., Veenstra J., Yi Q., McKinnon K., McKinnon J.E., Sexton J.Z., Zhou L, & Mi Q.S. (2022). Surface translocation of ACE2 and TMPRSS2 upon TLR4/7/8 activation is required for SARS-CoV-2 infection in circulating monocytes. Cell Discovery, 8, 89.

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