Nucleopore track etched membrane

Manufactured by Cytiva

Sourced in United Kingdom

Nucleopore track-etched membrane is a specialized filtration material used in various laboratory applications. It is produced by bombarding a thin polymer film with high-energy particles, creating uniform, precisely sized pores throughout the membrane. This process results in a membrane with a defined pore size distribution, enabling controlled filtration and separation of particles, cells, or molecules based on their size.

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Lab products found in correlation

1 2 dioleoyl sn glycero 3 phosphatidylcholine dopc, by Avanti Polar Lipids (1 mentions) Degassing station, by TA Instruments (1 mentions) Mini extruder, by Avanti Polar Lipids (1 mentions) L8 60m ultracentrifuge, by Beckman Coulter (1 mentions) Clear glass vials, by Thermo Fisher Scientific (1 mentions) Slide a lyzer dialysis cassette, by Thermo Fisher Scientific (1 mentions) Zeba spin desalting column, by Thermo Fisher Scientific (1 mentions) Dynapro plate reader 2, by Wyatt Technology (1 mentions) Cholera enterotoxin, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) Insulin, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Hydrocortisone, by Merck Group (1 mentions) Adenine, by Merck Group (1 mentions)

Close spelling variants of this product

Nucleopore (30 citations) Nucleopore membrane (17 citations) Nucleopore®-Track Etched (2 citations) Whatman® Nucleopore track-etched membranes (2 citations)

10 protocols using nucleopore track etched membrane

Microplastic Extraction and Quantification in Mosquitoes

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One individual was randomly removed from each beaker once all mosquitoes in the beaker had moulted into the 4 th instar, and again when they pupated or emerged as adults. All samples were then washed twice with distilled water to remove MPs from the surface of the mosquito and placed in separate 1.5 mL Eppendorf tubes, before being stored at -20 ºC prior to examination.
Microplastics were extracted from mosquitoes by homogenization and filtration. Mosquitoes were homogenized using a glass pestle in Eppendorf tubes containing 500 µL distilled water.
Individuals treated with 2 µm MPs were filtered through a nucleopore track-etched membrane (Whatman, UK) of < 1 µm and 25 mm dia.. Those exposed to 15 µm MPs were filtered through a nucleopore track-etched membrane (Whatman, UK) of < 10 µm and 25 mm dia. using a glass vacuum filter holder connected to a manual air pump. The MPs captured by both filters were quantified under a 20 × epi-fluorescent microscope (Zeiss Axioskop, USA). Adults were further dissected under a binocular stereo microscope (0.7 × -4.5 ×) to extract the gut and quantify the numbers of MPs under the epi-fluorescent microscope (Coleman et al., 2007) (link).

Al-Jaibachi R., Cuthbert R.N, & Callaghan A. (2019). Examining effects of ontogenic microplastic transference on Culex mosquito mortality and adult weight. The Science of the total environment, 651(Pt 1).

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DOPC Liposome Preparation Protocol

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1,2-Dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC; Avanti Polar Lipids) was dissolved in chloroform to prepare a 25 mg/ml stock (∼32 mM). The desired lipid amount (1.5 mg/ml, ∼2 mM) was transferred to a 15 ml Pyrex glass tube and dried under argon gas using a TurboVap evaporation system (Caliper). Lipid films were resuspended in pre-warmed volumes of 1× PBS buffer and rehydrated by incubation at 37°C for 2 h. Rehydrated liposomes were subjected to five freeze–thaw cycles using liquid nitrogen/warm water (40°C) followed by extrusion using a mini-extruder device (Avanti Polar Lipids) with 20 passages through polycarbonate membrane (pore size 0.1 μm, diam. 19 mm; Whatman Nucleopore Track-Etched membranes). The extruded liposomes (∼2 mM) were collected and transferred into sealed fresh glass tubes for further use.

Uppal S., Liu T., Galvan E., Gomez F., Tittley T., Poliakov E., Gentleman S, & Redmond T.M. (2022). An inducible amphipathic α-helix mediates subcellular targeting and membrane binding of RPE65. Life Science Alliance, 6(1), e202201546.

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Liposome Preparation via Extrusion

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The desired aqueous solution comprising ultrapure water (Milli-Q-water, 18.2 MΩ.cm, pH 4–6) was used to hydrate the lipids above their melting transition (55°C). The dispersion was softly stirred at 600 rpm for 30–40 min at 55°C using a Degassing Station (TA Instruments). This procedure yields multilamellar vesicles (LMV or MLV). LUV were prepared from suspension of MLV by extrusion through 100 nm polycarbonate membranes (Nucleopore Track-Etched Membranes, Whatman), above the melting transition of lipids using a Mini-Extruder (Avanti Polar Lipids). A lipid concentration of 4 mM was used in all the experiments.

Pérez-Isidoro R., Sierra-Valdez F.J, & Ruiz-Suárez J.C. (2014). Anesthetic Diffusion Through Lipid Membranes Depends on the Protonation Rate. Scientific Reports, 4, 7534.

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Encapsulation of Asparaginase in Liposomes

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Long-circulating liposomes were prepared by encapsulating the purified asparaginase (1 mg mL⁻¹) in their aqueous internal space. A mixture of the lipids Egg-PC:Chol:DSPE-PEG (68.25:30.5:1.25 molar ratio) at 32 mM was used. Liposomes were prepared by the dehydration-rehydration method, followed by extrusion, as previously described [9 (link),23 ,27 (link)]. The lipid dispersion containing the enzyme was sequentially extruded through membranes with decreasing pore diameter of 0.6, 0.4, 0.2, and 0.1 μm (Nucleopore^® Track-Etched Membranes, Whatman^®, Florham Park, NJ, USA) using Lipex Thermobarrel extruder. The non-encapsulated protein was separated by ultracentrifugation at 300,000× g for 120 min at 15 °C in a Beckman L8-60 M ultracentrifuge followed by Sephadex G-200 size exclusion chromatography using 10 mM sodium citrate buffer + 145 mM NaCl, pH 6.

Girão L.F., Carvalheiro M.C., Ferreira-Silva M., da Rocha S.L., Perales J., Martins M.B., Ferrara M.A., Bon E.P, & Corvo M.L. (2021). ASP-Enzymosomes with Saccharomyces cerevisiae Asparaginase II Expressed in Pichia pastoris: Formulation Design and In Vitro Studies of a Potential Antileukemic Drug. International Journal of Molecular Sciences, 22(20), 11120.

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Nanoscale Theranostic Delivery Platforms

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Three different NS, PEG₄₅-bl-PPS₂₀ MC, PEG₁₇-bl-PPS₃₀ PS, and PEG₄₅-bl-PPS₄₄ FM, were assembled and loaded with the lipophilic NIRF imaging agent ICG using the thin-film hydration method as previously described.^{65 (link)} Briefly, 8.6 mM of each block copolymer was dissolved in 150 μL dichloromethane within 1.8 mL clear glass vials (ThermoFisher Scientific). After desiccation to remove the solvent, the resulting thin films were hydrated in 1 mL of phosphate-buffered saline (PBS) or 1 mL of ICG solution (0.258 mM in PBS solution) under shaking at 1500 rpm overnight. The single-layer PS were obtained by extrusion multiple times through 0.2 μm and then 0.1 μm nucleopore track-etched membranes (Whatman). The ICG-loaded NS were purified from free ICG by Zeba Spin Desalting Columns (7K MWCO, ThermoFisher Scientific) equilibrated with PBS solution, and dialyzed against PBS using Slide-A-Lyzer Dialysis Cassettes (7K MWCO, ThermoFisher Scientific).

Yi S., Allen S.D., Liu Y.G., Ouyang B.Z., Li X., Augsornworawat P., Thorp E.B, & Scott E.A. (2016). Tailoring Nanostructure Morphology for Enhanced Targeting of Dendritic Cells in Atherosclerosis. ACS nano, 10(12), 11290-11303.

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Fluorescent Lipid Vesicle Preparation

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A fluorescent lipid mixture composed of 62 mol% 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 16 mol% 1,2-dimyristoyl-sn-glycero-3-phospho(1’-rac-glycerol) (sodium salt) (DMPG), 16 mol% cholesterol, 5 mol% 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy-(polyethylene glycol)-1000] (ammonium salt) (DMPE PEG 1000), and 1 mol% (DiI) (ThermoFisher, D282) was dried under nitrogen gas and resuspended in 10 mM HEPES in Hank’s Balanced Salt Solution (HBSS, components purchased from Sigma-Aldrich). The resulting lipid mixture was vortexed for 5 minutes and shaken at 500 rpm overnight before being hand-extruded through a Nucleopore track-etched membrane with pore sizes of 0.03 µm, 0.1 µm, 0.2 µm, or 0.4 µm (Whatman). Liposome sizes after extrusion were determined by dynamic light scattering (DLS) using a Wyatt DynaPro Plate Reader II and Dynamics V7 software (Supplemental Table S1 and Fig. S1). Ten repeats were measured for each condition and the average liposome size was reported along with the polydispersity.

Kuhn J., Smirnov A., Criss A, & Columbus L. (2019). Quantifying CEACAM targeted liposome delivery using imaging flow cytometry. Molecular pharmaceutics, 16(6), 2354-2363.

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Radiolabeling of Liposomal Nanocarriers

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The model liposome for the radiolabeling studies was TargoSphere^® [11 (link),12 (link)], an umbrella term coined for various lipid-based nanocarriers developed by Rodos Biotarget. The thin-film hydration method followed by extrusion [13 (link)] was used for preparing liposomes. In brief, phospholipids were dissolved, the stock solutions were combined in round-bottomed flasks, and lipid films were subsequently formulated by removing the solvents by means of a rotary evaporator. The resulting dry films were hydrated with PBS, and the crude samples were extruded through polycarbonate membranes with a pore size of 200 nm (Whatman^® Nucleopore™ Track-Etched Membrane), followed by extrusion through 50 nm. For radiolabeling, 10–100 µL liposomal aliquots of 30–35 µg/µL lipid concentration were added to [⁸⁹Zr]Zr(oxinate)₄.

Polyak A., Bankstahl J.P., Besecke K.F., Hozsa C., Triebert W., Pannem R.R., Manstein F., Borcholte T., Furch M., Zweigerdt R., Gieseler R.K., Bengel F.M, & Ross T.L. (2021). Simplified 89Zr-Labeling Protocol of Oxine (8-Hydroxyquinoline) Enabling Prolonged Tracking of Liposome-Based Nanomedicines and Cells. Pharmaceutics, 13(7), 1097.

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Opa Protein Folding and Reconstitution

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Opa protein folding was adapted from previously published protocols^{20 (link)–21 (link)}. 1,2-didecanoyl-sn-glycero-3-phosphocholine (diC10PC, Avanti Polar Lipids) dissolved in chloroform was dried under nitrogen gas and resuspended in borate buffer [10 mM sodium borate (pH 12.0) and 1 mM EDTA], then sonicated for 30 minutes at 40% amplitude (Q500, Q Sonica) in order to form liposomes. Following sonication, 4 M urea was added and 50 nm unfolded Opa₆₀ or Opa(HV−) was aliquoted and mixed. The Opa/diC10PC-liposome mixture was incubated for 4 days at 37°C, after which folding was confirmed by SDS-PAGE (Fig S5). Following Opa folding, diC10PC-proteoliposomes were pelleted through ultracentrifugation (142,400 x g for 2 hrs at 12°C), resuspended in resuspension buffer [10 mM HEPES (pH 7.4) in HBSS], and mixed with dried fluorescent lipids (DMPC, DMPG, cholesterol, DMPE-PEG-1000) as described above. The lipid mixture was vortexed for 5 minutes and shaken at 500 rpm for several hours before extrusion through a Nucleopore track-etched membrane with a 0.4 or 0.2 µm pore size (Whatman). Opa₆₀ reconstituted into liposomes are referred to as Opa₆₀ proteoliposomes.

Kuhn J., Smirnov A., Criss A, & Columbus L. (2019). Quantifying CEACAM targeted liposome delivery using imaging flow cytometry. Molecular pharmaceutics, 16(6), 2354-2363.

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Epidermal Explant Culture for Tissue Analysis

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A significant layer of mesoderm was left attached to the epidermis. Explants were immediately placed onto a solid support [PET-track-etched-membrane (Becton-Dickinson) or nucleopore-track-etched-membrane (Whatman)]. Explants cultured on PET-membranes were mounted on a sterile-grid support suspended on a Center-well organ culture dish (Falcon, 35307), overlain with a small amount of epidermal medium [DMEM (Invitrogen) supplemented with fetal bovine serum (10%), 1.8×10⁻⁴ M adenine (Sigma), 0.5 µg/ml hydrocortisone (Sigma), 5 µg/ml insulin (Sigma), 10⁻¹⁰ M cholera enterotoxin (Sigma), 10 ng/ml EGF (Peprotech)] and cultured at 37°C under 5% CO₂ (Jensen et al., 2010 (link)). Nucleopore membranes were floated on top of the epidermal medium and the explant covered by a drop of the same medium. Humidity was maintained using sterile 3 MM paper soaked in sterile PBS. Explants were cultured for 24 h and fixed in 4% formaldehyde in PBS prior to paraffin sectioning.

Panousopoulou E., Hobbs C., Mason I., Green J.B, & Formstone C.J. (2016). Epiboly generates the epidermal basal monolayer and spreads the nascent mammalian skin to enclose the embryonic body. Journal of Cell Science, 129(9), 1915-1927.

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Preparation of Large Unilamellar Vesicles

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Lipids, suspended in chloroform, were dried to form a thin film under N₂ while being vortexed in a clean glass tube. The lipid film was then placed under vacuum for 30 min to remove any residual solvent and then hydrated in either purified water or phosphate-buffered saline (PBS). In some cases, the resulting multilamellar vesicles (MLVs) were subjected to 10 freeze–thaw cycles. Large unilamellar vesicles (LUVs) were obtained by extruding the MLVs 31 times with a miniextruder (Avanti, 610000) using 10 mm–diameter filters (Whatman Drain Disc, 230300) and a membrane of pore size 0.1 µm (Whatman Nucleopore Track-Etched Membrane, 800309). The final concentration of lipid in LUV suspensions was either 3 or 1 mg/ml. LUVs were stored at 4°C and used within 2 wk.

Núñez M.F., Wisser K, & Veatch S.L. (2020). Synergistic factors control kinase–phosphatase organization in B-cells engaged with supported bilayers. Molecular Biology of the Cell, 31(7), 667-682.

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