Qev size exclusion column

Manufactured by Izon Science

Sourced in New Zealand, United Kingdom, Germany

The QEV size exclusion columns are laboratory equipment designed for the separation and purification of biological molecules based on their size. These columns utilize a porous gel media to separate molecules by allowing smaller ones to enter the pores, while larger molecules pass through more quickly. This size-based separation process is a core function of the QEV size exclusion columns.

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Lab products found in correlation

Amicon ultra 15 100 kda centrifugal filter units, by Merck Group (4 mentions) Amicon ultra 4 10 kda centrifugal filter, by Merck Group (3 mentions) Type 90 ti rotor, by Beckman Coulter (3 mentions) 0.2 μm filter, by Sarstedt (2 mentions) Vybrant did, by Thermo Fisher Scientific (2 mentions) Jem 1400 plus tem, by JEOL (2 mentions) Amicon ultrafiltration tube, by Merck Group (2 mentions) Amicon 10k centrifugation filters, by Merck Group (2 mentions) Bradford assay, by Bio-Rad (1 mentions) Exoquick ultra, by System Biosciences (1 mentions) Exoquick, by System Biosciences (1 mentions) Acs 1021, by American Type Culture Collection (1 mentions) Phenol red, by Lonza (1 mentions) Sypro ruby protein gel stain, by Bio-Rad (1 mentions) Cytofix, by BD (1 mentions) Bodipy fl, by Thermo Fisher Scientific (1 mentions) Ulbp1 pe, by R&D Systems (1 mentions) Fc receptor blocking reagent, by Miltenyi Biotec (1 mentions) Dmem glutamax media, by Thermo Fisher Scientific (1 mentions) H35 hypoxystation, by Don Whitley Scientific (1 mentions)

Close spelling variants of this product

QEV columns (48 citations) QEV Original Size Exclusion Chromatography (SEC) Columns (3 citations)

32 protocols using qev size exclusion column

Virus Purification via Size Exclusion

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For purification of virus samples, commercially available size exclusion columns (qEV, Izon Science, New Zealand) were used. The procedure was carried out as described by the manufacturer. In brief: 0.5–1 ml of clarified cell culture medium (centrifuged at 2200×g and filtered through 0.45-µm syringe filters) was overlaid on qEV size exclusion columns (Izon Science Ltd, New Zealand) followed by elution with PBS. 0.5 ml fractions were collected (and used immediately or stored at −80 °C before further analysis). The columns contain 10-ml resin material (with a pore size of approximately 75 nm) and has a void volume of 3 ml and a nominal separation size of 70 nm.

Heider S., Muzard J., Zaruba M, & Metzner C. (2017). Integrated Method for Purification and Single-Particle Characterization of Lentiviral Vector Systems by Size Exclusion Chromatography and Tunable Resistive Pulse Sensing. Molecular Biotechnology, 59(7), 251-259.

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Exosome Isolation via Size Exclusion

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Extracellular vesicles obtained from ultracentrifugation were loaded onto qEV size exclusion columns (Izon, Oxford, UK) for exosome isolation, as per manufacturer instructions. This technique allows the separation of particles from EV pellet based on their size into 16 fractions. The 16 fractions were concentrated using a vacuum centrifuge for 1.5 h at room temperature. Plasma exosomes of each animal were stored at −80 °C for exosome characterization and further analysis.

Almughlliq F.B., Koh Y.Q., Peiris H.N., Vaswani K., Holland O., Meier S., Roche J.R., Burke C.R., Crookenden M.A., Arachchige B.J., Reed S, & Mitchell M.D. (2019). Circulating exosomes may identify biomarkers for cows at risk for metabolic dysfunction. Scientific Reports, 9, 13879.

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Extracellular Vesicle Isolation by Size Exclusion

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EV were isolated using size exclusion column method as we described previously (16) . Briefly, conditioned media was collected and EV were isolated by centrifugation at 3000 rpm for 30 min to remove cells and debris, followed by filtration through a 0.22 μm filter to remove the remaining debris. Then the medium was further concentrated using Amicon Ultra-15 100 KDa centrifugal filter units (Millipore). Isolation of EV in the concentrated medium was carried out through qEV size exclusion columns (Izon Science). EV fractions were collected and concentrated by Amicon Ultra-4 10 KDa centrifugal filter (Millipore). The purified EV were stored at -80°C and subsequently characterized by particle size and electron microscopy.

Xuan W., Khan M., & Ashraf M. (2020). Human iPS Cells Derived Skeletal Muscle Progenitor Cells Promote Myoangiogenesis and Restore Dystrophin in Duchenne Muscular Dystrophic Mice.

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Isolation of NP Patient Exosomes from NLF

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Exosomes were isolated from NLF of NP patients (NPs exosomes) and healthy volunteers (normal exosomes) using a previously described method (28 (link),29 (link)) with the modifications that included differential centrifugation of NLF (6,000 × g for 30 min at 4°C and 10,000 × g for 60 min at 4°C) followed by ultrafiltration (0.2-µm filter; Sarstedt, Nümbrecht-Rommelsdorf, Germany) and qEV size-exclusion columns (iZON Science, Christchurch, New Zealand). The supernatant of NLF was then ultracentrifuged (100,000 × g) for 60 min at 4°C (Type 90 Ti Rotor; Beckman Coulter, Inc., Brea, CA, USA) to pellet the exosomes. The exosome pellets were then washed once with PBS.

Zhang W., Zhang J., Cheng L., Ni H., You B., Shan Y., Bao L., Wu D., Zhang T., Yue H, & Chen J. (2018). A disintegrin and metalloprotease 10-containing exosomes derived from nasal polyps promote angiogenesis and vascular permeability. Molecular Medicine Reports, 17(4), 5921-5927.

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Purification and Characterization of Extracellular Vesicles

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Five-hundred microlitres of clarified CCM, or 1 mL of processed plasma (centrifuged at 1,500 g and 10,000 g for 10 and 20 minutes, respectively) was overlaid on qEV size exclusion columns (Izon) followed by elution with PBS. Five-hundred-microlitre fractions were collected, and particle and protein concentrations determined with TRPS and the Bradford assay (Bio-Rad), respectively (Supplementary Fig. 3). High particle/low protein fractions from CCM were pooled and concentrated in Amicon^® Ultra-4 10 kDa nominal molecular weight centrifugal filter units to a final volume of 200 µL. Fractions from plasma were pooled and filtered with an Ultrafree^® 0.22 µm centrifugal filter device before being concentrated in an Amicon^® Ultra-4 10 kDa device.

Lobb R.J., Becker M., Wen Wen S., Wong C.S., Wiegmans A.P., Leimgruber A, & Möller A. (2015). Optimized exosome isolation protocol for cell culture supernatant and human plasma. Journal of Extracellular Vesicles, 4, 10.3402/jev.v4.27031.

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Exosome Isolation and Purification

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ExoQuick Ultra (System Biosciences, Palo Alto, CA) was added to the supernatant from the above step at a 1:4 ratio, mixed gently, and incubated for 30 min at 4 °C. After the incubation, the admixture was centrifuged at 1500 × g for 30 min to recover exosomes for total RNA isolation. For gel purification, serum was diluted with PBS (two-fold) and processed using qEV size exclusion columns (IZON, Medford, MA) according to manufacturer’s instructions.

Kumar S.R., Kimchi E.T., Manjunath Y., Gajagowni S., Stuckel A.J, & Kaifi J.T. (2020). RNA cargos in extracellular vesicles derived from blood serum in pancreas associated conditions. Scientific Reports, 10, 2800.

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Exosome Isolation and Characterization

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Exosomes were isolated by sequential ultracentrifugation(19 (link)). Briefly, CM harvested from cell culture was centrifuged at 300 × g for 10 minutes to remove cells. Exosomes were collected by centrifugation at 2,000 × g for 20 minutes. The supernatant was again centrifuged at 16,500 × g for 20 minutes to remove microvesicles. The supernatant was then passed through a 0.2μm filter (Sarstedt, Nümbrecht, GERMANY) to remove particles larger than 200 nm. Exosomes were then collected by ultracentrifugation at 120,000 × g for 70 minutes. Pellets were washed in PBS and centrifuged 70 minutes at 120,000 × g for 70 min and finally suspended in PBS.
Plasma exosomes were isolated as described previously(19 (link)). In brief, 1ml of serum was loaded onto the qEV size exclusion columns (Izon), and exosomes were elution with PBS. Isolated exosomes were concentrated by centrifugation using the Amicon® Ultra-4 10 kDa column.
The size distribution of isolated exosomes was analyzed by nano-particle tracking analysis using Nanosight NS50 and by eletronmicroscopy.

Xing F., Liu Y., Wu S.Y., Wu K., Sharma S., Mo Y.Y., Feng J., Sanders S., Jin G., Singh R., Vidi P.A., Tyagi A., Chan M.D., Ruiz J., Debinski W., Pasche B.C., Lo H.W., Metheny-Barlow L.J., D’Agostino RB J.r, & Watabe K. (2018). Loss of XIST in breast cancer activates MSN-c-Met and reprograms microglia via exosomal microRNA to promote brain metastasis. Cancer research, 78(15), 4316-4330.

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Exosome Isolation from Serum

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The exosomes were isolated from serum by precipitation or size exclusion chromatography (SEC). For precipitation, ExoQuick precipitation was carried out according to manufacturer’s instructions (System Biosciences). Briefly, one milliliter of serum was thawed and centrifuged at 1500g for 10 min. The supernatant was collected and recentrifuged at 10,000g for 30 min. The supernatant was then incubated with ExoQuick for 30 min at 4 °C. The ExoQuick/serum sample was then centrifuged at 1500g for 30 and the pellet was resuspended in 200 µL of PBS. For SEC, 500 µL of serum was thawed and recentrifuged at 10,000g for 30 min. The clarified serum was overlaid on qEV size exclusion columns (Izon, New Zealand) followed by elution with PBS. The eluate was collected in 13 sequential fractions of 1 mL. Each fraction was aliquoted and stored at − 80 °C for subsequently study.

Liu L., Pang H., He Q., Pan B., Sun X., Shan J., Wu L., Wu K., Yao X, & Guo Y. (2021). A novel strategy to identify candidate diagnostic and prognostic biomarkers for gastric cancer. Cancer Cell International, 21, 335.

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Isolation and Characterization of Extracellular Vesicles from Cardiac Progenitor Cells

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Human iPSC cell line ACS-1021 (ATCC, USA) and CPCs induced by ISX-9 were cultured as described [15 (link)]. In some cases, EBs and commercial human CPCs were also cultured. Conditioned media was collected and EVs were isolated by centrifugation at 3000 rpm for 30 min to remove cells and debris, followed by filtration through a 0.22 μm filter to remove the remaining debris. Then, the medium was further concentrated to 500 μl using Amicon Ultra-15 100 kDa centrifugal filter units (Millipore). Isolation of EV in the concentrated medium was carried out through qEV size exclusion columns (Izon Science). EV fractions were collected and concentrated by Amicon Ultra-4 10 kDa centrifugal filter units to a final volume of <100 μl. The purified EVs were stored at -80°C and subsequently characterized by particle size, EV markers, and electron microscopy.

Xuan W., Wang L., Xu M., Weintraub N.L, & Ashraf M. (2019). miRNAs in Extracellular Vesicles from iPS-Derived Cardiac Progenitor Cells Effectively Reduce Fibrosis and Promote Angiogenesis in Infarcted Heart. Stem Cells International, 2019, 3726392.

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Fractionation of Raw Cow Milk Proteins

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To fractionate raw cow’s milk based on molecular size, qEV size exclusion columns (Izon Science, Oxford, UK) were used according to the manufacturer’s protocol. Briefly, 0.5 mL of 10,000× g raw milk supernatant (free of cells, cell debris, and cream) was loaded onto the size exclusion column (Izon Science) and the first 3 mL of eluent was discarded. Eluent fractions of 0.5 mL were then collected up to 12 mL (24 fractions), by continuously adding RPMI 1640 medium without l-glutamine and phenol red (Lonza) to the column. Protein content of each fraction was quantified by using a NanoDrop ND-1000 spectrophotometer (A280; Thermo Fisher Scientific). To determine the molecular weight of the proteins in each fraction, proteins were separated by using a 12.5% SDS-PAGE under non-reducing conditions and visualized with SYPRO^® Ruby Protein Gel Stain (Bio-Rad, Veenendaal, The Netherlands). Fractions were stored at −80 °C until further use.

Abbring S., Blokhuis B.R., Miltenburg J.L., Olmedo K.G., Garssen J., Redegeld F.A, & van Esch B.C. (2020). Direct Inhibition of the Allergic Effector Response by Raw Cow’s Milk—An Extensive In Vitro Assessment. Cells, 9(5), 1258.

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