Ev depleted fbs

Manufactured by Thermo Fisher Scientific

Sourced in United States

EV-depleted FBS is a cell culture supplement that has been processed to remove extracellular vesicles (EVs) from the fetal bovine serum (FBS). This product is intended to provide a serum source for in vitro cell culture while minimizing potential interference from EVs.

Automatically generated - may contain errors

Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Bradford method, by Bio-Rad (1 mentions) Fdls3000, by Shimadzu (1 mentions) Jem 1400flash, by JEOL (1 mentions) Milli q system, by Merck Group (1 mentions) Nitrocellulose membrane, by Cytiva (1 mentions) Gf 1 nucleic acid extraction kit, by Vivantis Technologies (1 mentions) Duoset elisa development system, by R&D Systems (1 mentions) L glutamine, by Merck Group (1 mentions) Hepes, by Merck Group (1 mentions) Hemin, by Merck Group (1 mentions) Adenosine, by Merck Group (1 mentions) Gentamicin, by Merck Group (1 mentions) Bca protein assay kit, by R&D Systems (1 mentions) Beta actin monoclonal antibody, by Proteintech (1 mentions) Nanosight ns300, by Malvern Panalytical (1 mentions) Nta software, by Malvern Panalytical (1 mentions) Lps from e coli serotype 055 b5, by Merck Group (1 mentions) Polypropylene tube, by Greiner (1 mentions) Dmem glutamax, by Thermo Fisher Scientific (1 mentions)

16 protocols using ev depleted fbs

Isolation and Characterization of Extracellular Vesicles

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

The medium of subconfluent AT-MSCs or WJ-MSCs was changed to Iscove's modified Dulbecco's medium (IMDM) with 1% penicillin/streptomycin and 0.25% EV-depleted FBS (Thermo Fisher Scientific). After 24 h, the supernatants were collected and centrifuged at 1000 rpm for 5 min, and then centrifuged at 2100 rpm for 20 min to remove cell debris from the medium. Cell-free supernatants were ultracentrifuged at 100,000 rpm for 70 min at 4°C, and the pellets were stained with PKH26 (PKH26 linker; Sigma-Aldrich, St. Louis, MO) for 5 min at room temperature. After that, the washing step was performed twice by adding PBS with 0.25% EV-depleted FBS (Thermo Fisher Scientific) and ultracentrifuged at the same conditions. After washing, pellets of EVs were collected.
EVs then were characterized by quantification of the protein concentration using the Bradford method (Bio-Rad, Hercules, CA), size measurement using a particle size analyzer (FDLS3000; Shimadzu Corporation, Kyoto, Japan), and a morphology analysis by transmission electron microscopy (JEM-1400Flash; JEOL Ltd., Tokyo, Japan).

Khanh V.C., Fukushige M., Chang Y.H., Hoang N.N., Yamashita T., Obata-Yasuoka M., Hamada H., Osaka M., Hiramatsu Y, & Ohneda O. (2021). Wharton's Jelly Mesenchymal Stem Cell-Derived Extracellular Vesicles Reduce SARS-CoV2-Induced Inflammatory Cytokines Under High Glucose and Uremic Toxin Conditions. Stem Cells and Development, 30(15), 758-772.

+ Open protocol

+ Expand

Chondrogenic Differentiation of ADSCs

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

ADSCs were seeded in 12-well plates at a density of 5000 cells/well and the upper chamber of 8.0-µm Transwell plates (Corning) at a high density of 8 × 10⁴ cells. Cells were cultured overnight in a medium supplemented with 10% FBS and then rinsed twice with DPBS after removing the culture medium. The replacement culture medium of the upper chambers was culture medium containing 10% EV-depleted FBS, and the lower chambers were replaced with chondrogenic differentiation medium containing 10% EV-depleted FBS. The medium was refreshed every 3 days. On day 21, the efficiency of cartilage differentiation was assessed by histochemical staining (Alcian blue). Cells were analyzed under a light microscope (Olympus BX43, Tokyo, Japan). Commercially available EV-depleted FBS was purchased from Gibco (Life Technologies, Waltham, MA, USA).

Tsai Y.C., Cheng T.S., Liao H.J., Chuang M.H., Chen H.T., Chen C.H., Zhang K.L., Chang C.H., Lin P.C, & Huang C.Y. (2022). Mesenchymal Stem Cell Secreted-Extracellular Vesicles are Involved in Chondrocyte Production and Reduce Adipogenesis during Stem Cell Differentiation. Tissue Engineering and Regenerative Medicine, 19(6), 1295-1310.

+ Open protocol

+ Expand

Real-Time PCR and Western Blot Protocols

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

All solutions were prepared by apyrogenic deionized water (MilliQ System, Millipore, France). All materials used for real-time PCR were purchased from Qiagen (Germany), except the DNA extraction kit that was GF-1 Nucleic Acid Extraction Kit and was purchased from Vivantis Technologies (Malaysia). The GP63 antibody was provided by the Pasteur Institute of Iran, the anti-GFP-HRP goat polyclonal antibody was obtained from Acris antibodies GmbH (Germany), and the beta actin monoclonal antibody was purchased from ProteinTech Group Inc. (Chicago, IL, USA). The nitrocellulose membrane was supplied from Amersham (UK), and the ECL Western blotting substrate was from Pierce™ (Pierce Biotechnology, Thermo Fisher, Rockford, IL, USA). The bovine serum albumin (BSA), phorbol 12-myristate 13-acetate (PMA), Resazurin powder, M199 and RPMI-1640 cell culture media, HEPES, L-glutamine, adenosine, gentamicin, and hemin were sourced from Sigma (Germany). The fetal bovine serum (FBS) and EV-depleted FBS were obtained from Gibco (Life Technologies, Germany). The bicinchoninic acid (BCA) and ELISA kits were purchased from Pierce™ (BCA Protein Assay Kit, USA) and DuoSet ELISA development system (R&D Systems, USA), respectively.

Shokouhy M., Sarvnaz H., Taslimi Y., Lajevardi M.S., Habibzadeh S., Mizbani A., Shekari F., Behbahani M., Torrecilhas A.C, & Rafati S. (2022). Isolation, characterization, and functional study of extracellular vesicles derived from Leishmania tarentolae. Frontiers in Cellular and Infection Microbiology, 12, 921410.

+ Open protocol

+ Expand

Extracellular Vesicle Isolation from Cell Lines

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

Cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and confirmed to be free of mycoplasma contamination [24 (link)]. All cell lines were cultured in a humidified environment at 37 °C, 5% CO₂ and seeded to 70% confluence for 24 h. BEAS-2B cells were cultured in LHC-9 medium, A549 cells in DMEM plus 10% FBS, and the remaining lines in RPMI plus 10% FBS (Gibco/ThermoFisher Scientific, Waltham, MA, USA). Prior to EV collection, cells were washed with DPBS and incubated in their respective media for 48 h and using EV-depleted FBS (Gibco/ThermoFisher Scientific) as required. Conditioned media were collected and processed for EV isolation. To study LINE-1 inducible conditions, 2 × 10⁶ H460 cells were seeded in 150-mm dishes and allowed to attach overnight. Cells were washed with DPBS and incubated with RPMI medium containing 10% EV-depleted FBS plus 1 μM BaP dissolved in 0.2% DMSO. Conditioned media were collected after 48 h and processed for EV isolation.

Bowers E.C., Motta A., Knox K., McKay B.S, & Ramos K.S. (2022). LINE-1 Cargo and Reverse Transcriptase Activity Profiles in Extracellular Vesicles from Lung Cancer Cells and Human Plasma. International Journal of Molecular Sciences, 23(7), 3461.

+ Open protocol

+ Expand

Extracellular Vesicle Isolation and Characterization

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

Twenty-four hours after transfection, the cell culture medium was replaced with DMEM that contains EV-depleted FBS (Gibco). Seventy-two hours after transfection, cell culture supernatants were precleared via two consecutive rounds of centrifugation (500g for 10 min and 2000g for 10 min) and filtered using a 0.2-μm syringe filter (Pall Acrodisc, catalog no. 4562). EVs in the supernatants were then pelleted by ultracentrifugation using SW32 Ti rotor at 100,000g for 2 hours. EV pellet was resuspended in filtered PBS and used immediately or stored at 4°C. EVs were analyzed and quantified by the NanoSight NS300 instrument with NTA software (Malvern Panalytical).

Choi S., Yang Z., Wang Q., Qiao Z., Sun M., Wiggins J., Xiang S.H, & Lu Q. (2023). Displaying and delivering viral membrane antigens via WW domain–activated extracellular vesicles. Science Advances, 9(4), eade2708.

+ Open protocol

+ Expand

EV Production from LPS-Stimulated THP-1 Cells

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

For EV production and harvest, THP-1 cells were collected in 50 ml polypropylene tubes (Greiner Bio One, Frickenhausen, Germany, #227261) by 10 minutes of centrifugation at 200 rcf and 20°C in a swinging bucket Rotanta 460 R centrifuge (Hettich, Tuttlingen, Germany). Next, they were seeded at a density of 1×10⁶ cells/ml in a T75 cm² tissue culture treated cell culture flask (Greiner Bio One, #658170) in 15 ml of RPMI 1460 medium with GlutaMAX supplemented with 2% EV-depleted FBS (Gibco, #A2720801), and stimulated with 100 ng/ml lipopolysaccharide (LPS) from E. coli serotype 055:B5 (Sigma-Aldrich, St. Louis, USA, #L2880). After 24 hours of incubation at 37°C, 5% CO₂ and 95% humidity, cells and the conditioned cell medium were collected.

Deville S., Berckmans P., Van Hoof R., Lambrichts I., Salvati A, & Nelissen I. (2021). Comparison of extracellular vesicle isolation and storage methods using high-sensitivity flow cytometry. PLoS ONE, 16(2), e0245835.

+ Open protocol

+ Expand

Extracellular Vesicle Isolation and Characterization

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

Cells were seeded either in 12-well plates (250 000 cells/well) for immunoblotting and RT-PCR analyses or in T75 flasks (2 million cells/flask) for immunoblotting analyses and EV isolation. The cells were cultured in DMEM-Glutamax (Gibco), supplemented with 50 U/mL Penicillin, 50 U/mL Streptomycin (Gibco), and either 10% FBS (PAA Laboratories) or 10% EV-depleted FBS (Gibco). For stimulation, cells were exposed to 2.5 µM oAβ, with or without EV inhibitors (20 µM GW4869 or 0.25 µM Manumycin A) dissolved in equal amounts of dimethyl sulfoxide (DMSO), for 6 h. At this point, medium and cells were either isolated or washed with PBS followed by an additional 48 h incubation in 25 mL cell medium for flasks and 2.5 mL for 12-well plates, with or without EV inhibitors. The control medium contained the same amount of DMSO as the EV-inhibitor medium (0.58%). After 48 h, cells were collected for immunoblotting purposes and frozen until further use. Cell medium was collected for EV isolation and centrifuged at 2000xg for 10 min using a TX-400 swinging bucket rotor in a Heraeus Multifuge X1R (Thermo Fisher Scientific) centrifuge to obtain the supernatant, which was frozen until further use.

Johansson L., Reyes J.F., Ali T., Schätzl H., Gilch S, & Hallbeck M. (2024). Lack of cellular prion protein causes Amyloid β accumulation, increased extracellular vesicle abundance, and changes to exosome biogenesis proteins. Molecular and cellular biochemistry.

+ Open protocol

+ Expand

EV-Depletion and LPS Treatment Assay

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

LPSs from Escherichia coli O111:B4, PMA, and PEG 6000 were purchased from Sigma-Aldrich. PBS, TBS, penicillin-streptomycin (10,000 U/mL), FBS, EV-depleted FBS, DMEM, and RPMI-1640 were purchased from Gibco. Lipofectamine 2000 reagent, protease inhibitor tablet, and BSA were purchased from Thermo Fisher Scientific. The K. pneumoniae strain was purchased from ATCC (#43186, Manassas, VA). Recombinant human CC16 was purchased from Novus Biologicals (NBP2-35126, Centennial, CO).

Han Y., Zhu Y., Almuntashiri S., Wang X., Somanath P.R., Owen C.A, & Zhang D. (2023). Extracellular vesicle-encapsulated CC16 as novel nanotherapeutics for treatment of acute lung injury. Molecular Therapy, 31(5), 1346-1364.

+ Open protocol

+ Expand

Murine MSC Extracellular Vesicle Isolation

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

Murine MSCs were seeded 1 × 10⁶ cells per 10 cm² and incubated overnight. Following the overnight incubation, cells were washed in phosphate-buffered saline (PBS) and fresh media containing EV-depleted FBS (Gibco, UK) was applied. EV-depleted FBS was obtained by centrifugation at 120,000×g for 18 h at 4 °C. Cells were incubated for 48 h under standard conditions. After 48 h, the media was collected. The conditioned medium was centrifuged at 400×g for 5 min at 4 °C. The resulting supernatant was sequentially centrifuged at 2,000×g for 20 min at 4 °C and 10,000×g for 45 min. Then the supernatant was transferred to ultracentrifuge tube and centrifuged at 100,000×g for 90 min at 4 °C using SW28Ti rotor (Beckman Coulter, USA) in the BECKMAN L70 ultracentrifuge (Beckman Coulter, USA).

Gomzikova M.O., Aimaletdinov A.M., Bondar O.V., Starostina I.G., Gorshkova N.V., Neustroeva O.A., Kletukhina S.K., Kurbangaleeva S.V., Vorobev V.V., Garanina E.E., Persson J.L., Jeyapalan J., Mongan N.P., Khaiboullina S.F, & Rizvanov A.A. (2020). Immunosuppressive properties of cytochalasin B-induced membrane vesicles of mesenchymal stem cells: comparing with extracellular vesicles derived from mesenchymal stem cells. Scientific Reports, 10, 10740.

+ Open protocol

+ Expand

HIV-1 NL4.3 Virus Production and Characterization

Cited 2 times

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

TZM-bl cells (NIH AIDS Reagent Program, Germantown, MD), and 293T cells (ATCC) were maintained in complete DMEM (Gibco-BRL/Life Technologies, Carlsbad, CA) containing 5% EV-depleted FBS (Gibco, Carlsbad, CA) as described¹⁷. HIV-1 pNL4.3 plasmid (NIH AIDS Reagent Program) was used to generate HIV-1 NL4.3 virus in 293T cells^{4 (link)–6 (link)}. Virus titers were determined by TZM-bl renilla luciferase units (RLU) and EnzChek Reverse Transcriptase Assay (Life Technologies). Cell viability was determined by MTT assay with three replicates per donor at the time of infection analysis as described^{4 (link),17}.

Welch J.L., Kaddour H., Winchester L., Fletcher C.V., Stapleton J.T, & Okeoma C.M. (2020). Semen extracellular vesicles from HIV-1 infected individuals inhibit HIV-1 replication in vitro, and extracellular vesicles carry antiretroviral drugs in vivo. Journal of acquired immune deficiency syndromes (1999), 83(1), 90-98.

+ 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!