0.22 μm pore pes filter

Manufactured by Merck Group

Sourced in United States

The 0.22-μm pore PES filters are a type of laboratory equipment used for filtration. They feature a polyethersulfone (PES) membrane with a pore size of 0.22 microns, which is designed to remove particles, microorganisms, and other contaminants from liquids.

Automatically generated - may contain errors

Lab products found in correlation

Exosome free fbs, by System Biosciences (2 mentions) Malvern zetasizer nano zs90, by Malvern Panalytical (1 mentions) Cm120 biotwin transmission electron microscope, by Thermo Fisher Scientific (1 mentions) Advanced rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Glutamine, by Thermo Fisher Scientific (1 mentions) 100 kda cut off filter, by Merck Group (1 mentions) Sw41ti rotor, by Beckman Coulter (1 mentions) D pbs, by Nacalai Tesque (1 mentions)

4 protocols using 0.22 μm pore pes filter

Exosome Isolation and Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

For exosomes isolation, supernatant collected from ovarian cancer cells cultured in DMEM containing 10% Exosome-free FBS (System Biosciences, USA) for 48 h was centrifuged at 300 × g for 10 min, 3000 × g for 30 min and 10,000 × g for 30 min. Then, the supernatant was passed through 0.22-μm pore PES filters (Millipore). Ultracentrifugation was performed at 100,000 × g for 70 min using an Optima XE-90 Supercentrifuge (Beckman, Germany) to enrich the exosomes, and the pellet was rinsed using 30 ml of PBS. Finally, the exosomes were collected by ultracentrifugation at 100,000 × g for 70 min.
Isolated exosomes were mixed with 4% paraformaldehyde. exosomes were then dropped onto formvar carbon-coated electron microscopy grids and fixed with 1% glutaraldehyde for 10 min. Samples were negatively stained with 2% uranyl acetate solution. Images were obtained using Philips CM120 BioTwin transmission electron microscope (TEM) (FEI Company, USA).
Nanoparticle tracking analysis (NTA) was performed by Malvern Zetasizer Nano ZS-90 (Malvern Instruments Ltd., UK) following the manufacturer’s instructions. The exosomes were diluted in PBS. The mean particle size and size distribution were analyzed by dynamic light scattering method using the Malvern Zetasizer Nano ZS-90.

Li H., Zeng C., Shu C., Cao Y., Shao W., Zhang M., Cao H, & Zhao S. (2022). Laminins in tumor-derived exosomes upregulated by ETS1 reprogram omental macrophages to promote omental metastasis of ovarian cancer. Cell Death & Disease, 13(12), 1028.

+ Open protocol

+ Expand

Extracellular Vesicle Isolation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Growth medium used for EV collection was prepared with 10% FBS and ultracentrifuged at 110,000 × g for 18 h at 4°C to remove EVs in FBS. After 48 h of culture, EVs from cell‐conditioned medium were extracted by a standard differential centrifugation protocol. In brief, cell‐conditioned medium was serially centrifuged at 300 × g for 20 min, 3000 × g for 20 min and 10,000 × g for 60 min at 4°C to remove cells, debris and large vesicles. The supernatant was passed through 0.22 μm pore PES filters (Millipore) and then centrifuged at 110,000 × g for 70 min at 4°C to pellet EVs. Subsequently, EVs were washed once and finally resuspended using PBS.

Liu B., Jin Y., Yang J., Han Y., Shan H., Qiu M., Zhao X., Liu A., Jin Y, & Yin Y. (2022). Extracellular vesicles from lung tissue drive bone marrow neutrophil recruitment in inflammation. Journal of Extracellular Vesicles, 11(5), e12223.

+ Open protocol

+ Expand

Exosome Isolation from Cell Culture Supernatant

Check if the same lab product or an alternative is used in the 5 most similar protocols

For exosome isolation, MFF-1 cells grown in 150 mm cell culture dishes were washed thrice with PBS at approximately 80% confluence and cultivated with DMEM containing 10% FBS depleted bovine serum extracellular vehicles (EVs) by overnight ultracentrifugation at 100,000× g at 4 °C after transfection with plasmids. After 48 h, the conditioned medium (CM) was collected and first pre-cleared by centrifugation at 300× g for 10 min at RT to remove the floating cells. Additionally, all subsequent centrifugation steps were performed at 4 °C. Next, the supernatant was spun at 20,000× g for 20 min to remove dead cells and shedding vesicles. Then, to collect exosomes, the supernatant was isolated by ultracentrifugation at 110,000× g for 70 min (Ti70, Beckman-Coulter, Inc., Brea, CA, USA) and the supernatant was removed. Furthermore, the precipitate was resuspended in a large volume of PBS and passed through a 0.22 μm pore PES filter (Merck Millipore, Billerica, MA, USA). Then, this supernatant (pre-cleared medium) was next followed by ultracentrifugated at 110,000× g for 70 min (SW40, Beckman-Coulter, Inc., Brea, CA, USA) to sediment the exosomes. The precipitate was resuspended with PBS and subjected to the same ultracentrifugation conditions to wash the sample.

He J., Chen N.N., Li Z.M., Wang Y.Y., Weng S.P., Guo C.J, & He J.G. (2021). Evidence for a Novel Antiviral Mechanism of Teleost Fish: Serum-Derived Exosomes Inhibit Virus Replication through Incorporating Mx1 Protein. International Journal of Molecular Sciences, 22(19), 10346.

+ Open protocol

+ Expand

Isolation and Purification of Small Extracellular Vesicles

Check if the same lab product or an alternative is used in the 5 most similar protocols

For isolation of sEVs, 6.0 × 10⁶ PANC-1 cells were seeded in thirteen 150 mm dishes and precultured with RPMI 1640 containing FBS and antibiotics for 24 h. They were washed twice with Dulbecco’s phosphate-buffered saline (D-PBS, Nacalai Tesque). As FBS also includes large amounts of sEVs, it is necessary to change to advanced RPMI 1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 2 mmol/L glutamine (Thermo Fisher Scientific) and antibiotics. Thereafter, the cells were cultured for 48 h under hypoxia (1% O₂) or normal oxygen conditions (n = 3). Fresh (non-cultured) advanced RPMI 1640 medium was used as a blank (n = 3). Cell-conditioned medium was collected and centrifuged at 2000× g for 25 min at 4 °C to pellet and remove cells, debris, and apoptotic bodies. The supernatant was filtered using a 0.22 μm pore PES filter (Merck Millipore, Burlington, MA, USA) to remove large sEVs. The filtrate was concentrated with a 100 kDa cut-off filter (Merck Millipore). The concentrate was ultracentrifuged at 37,000 rpm (average RCF is 234,700× g) for 70 min at 4 °C (SW41Ti rotor, Beckman Coulter, Brea, CA, USA). The pellet was washed with physiological saline (Hikari Pharmaceutical, Tokyo, Japan) and was collected by ultracentrifuge. This washing procedure was repeated twice. Finally, the weight of the pellet was adjusted to 0.05 g (≈ 50 μL) by adding physiological saline.

Hayasaka R., Tabata S., Hasebe M., Ikeda S., Ohnuma S., Mori M., Soga T., Tomita M, & Hirayama A. (2021). Metabolomic Analysis of Small Extracellular Vesicles Derived from Pancreatic Cancer Cells Cultured under Normoxia and Hypoxia. Metabolites, 11(4), 215.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!