Mini extruder set

Manufactured by Avanti Polar Lipids

Sourced in United States

The Mini-extruder set is a laboratory instrument used for the extrusion of small-scale lipid samples. It facilitates the preparation of uniform lipid vesicles or liposomes through a controlled extrusion process. The core function of this equipment is to create homogeneous lipid dispersions by forcing the sample through a defined pore size membrane under gentle mechanical pressure.

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Lab products found in correlation

1 1 dioctadecyl 3 3 3 3 tetramethylindocarbocyanine perchlorate dii, by Thermo Fisher Scientific (2 mentions) 1 palmitoyl 2 oleoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (2 mentions) 1 2 dioleoyl sn glycero 3 phosphoethanolamine n cap biotinyl sodium salt, by Avanti Polar Lipids (2 mentions) Pc membrane, by Cytiva (2 mentions) Total gangliosides ga, by Avanti Polar Lipids (2 mentions) Polycarbonate membranes, by GE Healthcare (1 mentions) Zetasizer nano s instrument, by Malvern Panalytical (1 mentions) 100 nm polycarbonate membrane filter, by Cytiva (1 mentions) Pd 10 column, by GE Healthcare (1 mentions) Nuclepore, by Cytiva (1 mentions) E coli polar lipid extract, by Avanti Polar Lipids (1 mentions) Acquity arc system, by Waters Corporation (1 mentions)

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Avanti Mini-Extruder Set Extruder set Mini-extruder

13 protocols using mini extruder set

Transfersome Deformability Characterization

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In total, 1 mL of the transfersome suspension was extruded using a polycarbonate membrane with a pore size of 100 nm in a mini-extruder set (Avanti Polar Lipids Inc., USA). The extruded suspension volume in 5 min was recorded, and the particle size was then determined using the dynamic light scattering method. The deformability index was calculated using the following equation:

D = J {(\frac{r v}{r p})}^{2},

where D is the deformability index, J is the amount of transfersome suspension that passed through the membrane in 5 min (mL), rv is the particle size of the transfersomes that passed through the membrane (nm), and rp is the membrane pore size (nm).^{21 (link)}

Surini S., Leonyza A, & Suh C.W. (2020). Formulation and In Vitro Penetration Study of Recombinant Human Epidermal Growth Factor-Loaded Transfersomal Emulgel. Advanced Pharmaceutical Bulletin, 10(4), 586-594.

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Encapsulation of Nanoparticles in Liposomes

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NP-liposomes were prepared using a reverse-phase evaporation method^{40 (link)} to ensure maximum encapsulation of NPs into liposomes. One hundred μL of DNA-NPs (3 nM) were combined with 200 μL of a 12.5 mg mL ⁻¹ lipid solution in chloroform. The lipid mixture contained 98% (w/w) 1-palmitoyl-2-oleoyl-sn-gly-cero-3-phosphocholine (POPC) and 2% (w/w) 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) sodium salt (Avanti Polar Lipids Inc., Alabaster, AL, USA). For the colocalization experiments, the lipid mix also contained 2% (w/w) 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI; Invitrogen, Carlsbad, CA, USA), and the POPC content was reduced to 96%. The two-phase mixture was sonicated with a probe sonicator for 30 s. Then, chloroform was removed in vacuum on a rotary evaporator. The resulting viscous solution was diluted with P50 buffer and extruded through a 200-nm pore size polystyrene membrane with a mini extruder set (Avanti Polar Lipids Inc.). After the extrusion, the liposome solution was centrifuged at 100 g for 30 min to collect the formed NP-containing liposomes.

Chen T., Wang X., Alizadeh M.H, & Reinhard B.M. (2017). Monitoring transient nanoparticle interactions with liposome-confined plasmonic transducers. Microsystems & Nanoengineering, 3, 16086.

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Liposome Encapsulation of DNA Nanoparticles

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NP-liposomes were prepared using a reverse-phase evaporation method^{40 (link)} to ensure maximum encapsulation of NPs into liposomes. One hundred μL of DNA-NPs (3 nM) were combined with 200 μL of a 12.5 mg mL⁻¹ lipid solution in chloroform. The lipid mixture contained 98% (w/w) 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 2% (w/w) 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) sodium salt (Avanti Polar Lipids Inc., Alabaster, AL, USA). For the colocalization experiments, the lipid mix also contained 2% (w/w) 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI; Invitrogen, Carlsbad, CA, USA), and the POPC content was reduced to 96%. The two-phase mixture was sonicated with a probe sonicator for 30 s. Then, chloroform was removed in vacuum on a rotary evaporator. The resulting viscous solution was diluted with P50 buffer and extruded through a 200-nm pore size polystyrene membrane with a mini extruder set (Avanti Polar Lipids Inc.). After the extrusion, the liposome solution was centrifuged at 100 g for 30 min to collect the formed NP-containing liposomes.

Chen T., Wang X., Alizadeh M.H, & Reinhard B.M. (2017). Monitoring transient nanoparticle interactions with liposome-confined plasmonic transducers. Microsystems & Nanoengineering, 3, 16086.

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Preparation of EggPC Liposomes

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Small unilamellar liposomes were prepared by 1.3 mg of EggPC dissolved in chloroform, followed by drying in a round-bottom flask in vacuo. The dried thin film of EggPC was hydrated in a 1 mL of 100 mM phosphate buffer solutions (containing 100 mM sodium chloride) with different pH values. The resultant the multilamellar vesicle was subjected to 5 freeze-thaw cycles. Then, the solution of heterogeneous size of unilamellar liposomes is passed through a polycarbonate filter with 100 nm diameter pores (Merck, Darmstadt, Germany) a total of 21 times on Mini-Extruder Set (Avanti Polar Lipids, Inc., Alabaster, AL, USA).
Liposomes consisting of 2.0 mM EggPC and incorporated with varying amounts (0.5, 1.0, 1.5 and 2.5 mol%) of the rLPE-St were also prepared by the same procedure described above.

Kashiwada A., Namiki K., & Mori H. (2020). Design and Construction of pH-Selective Self-Lytic Liposome System. Processes, 8(12), 1526-1526.

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Liposomal 5-FU Delivery Protocol

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The liposomes were prepared by film hydration method followed by sequential extrusion. Briefly, PC, CHOL, and DSPE-PEG-MAL, in different molar ratio (Table 1), were dissolved in chloroform in a round-bottom flask (250 mL). After dissolution, the solvent was evaporated under reduced pressure, at room temperature, in a rotary evaporator—Hei-VAP ML from Heidolph (Schwabach, Germany)—at 160 rpm, leading to the formation of a thin and homogeneous film of lipids on the surface of the flask.
The film obtained was then hydrated using 5 mL of 5-FU aqueous solution in phosphate-buffered saline (pH = 7.4) (10 or 15 mg/mL) at room temperature. After the vortex shaking, a spontaneous formation of multilamellar vesicles occurred. The dispersion was placed in a bath sonicator for 30 min at 25 °C in order to break the existing aggregates. Finally, the size of liposomes was reduced by multiple extrusion steps (10 times) through polycarbonate membranes with a pore size of 0.2 μm using a mini-extruder set from Avanti Polar Lipids, Inc (Alabaster, AL, USA). Free 5-FU was removed by dialysis method using a cellulose tubular membrane (12000–14000 Da).

Cadinoiu A.N., Rata D.M., Atanase L.I., Daraba O.M., Gherghel D., Vochita G, & Popa M. (2019). Aptamer-Functionalized Liposomes as a Potential Treatment for Basal Cell Carcinoma. Polymers, 11(9), 1515.

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Preparation of DMPC Unilamellar Liposomes

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Powdered DMPC was dissolved in chloroform to a final lipid concentration of 7.37 mM and if desired mixed with 1 mol% of lipid spin label. Chloroform was evaporated under a stream of nitrogen gas. The resulting lipid film was dried under vacuum for at least 2 h. The dried lipids were suspended in 50 mM Tris (pH 7.5), 200 mM NaCl (buffer D) and vortexed. Subsequently, the multilamellar liposome suspension underwent five freeze–thaw cycles (N₂/water bath at 37 °C), and, if not used directly, was stored in aliquots at −80 °C. Before reconstitution, the suspension of liposomes was extruded 31 times through polycarbonate membranes of 100 nm pore size using a Mini-Extruder Set (Avanti Polar Lipids, Alabaster, AL, USA) to get unilamellar vesicles.

Voskoboynikova N., Orekhov P., Bozdaganyan M., Kodde F., Rademacher M., Schowe M., Budke-Gieseking A., Brickwedde B., Psathaki O.E., Mulkidjanian A.Y., Cosentino K., Shaitan K.V, & Steinhoff H.J. (2021). Lipid Dynamics in Diisobutylene-Maleic Acid (DIBMA) Lipid Particles in Presence of Sensory Rhodopsin II. International Journal of Molecular Sciences, 22(5), 2548.

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Preparation of Functionalized Lipid Vesicles

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Lyophilised 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) was dissolved in chloroform, with or without head group-functionalized lipid (i.e. 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(Cap biotinyl) (referred to here as BC-PE) or 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[3-(2-pyridyldithio)propionate] (termed PDP-PE)) (Avanti Polar Lipids, USA), within a glass tube. Chloroform was removed under a continuous nitrogen stream to produce a thin lipid film. Tubes were placed under vacuum for a minimum of 2 h to ensure all chloroform had been removed. Lipid films were rehydrated in 20 mM HEPES, 150 mM NaCl, 2 mM CaCl₂, pH 7.4 at 1 mg ml⁻¹. Lipid solutions were extruded through a 50 nm pore size filter composed of polycarbonate membranes (Whatman®, GE Healthcare Life Sciences), using a mini-extruder set (Avanti Polar-Lipids, USA). Dynamic Light Scattering (DLS) using a Zetasizer nano-S instrument (Malvern Instruments Ltd, UK) was used to check the size of the unilamellar vesicles. Only vesicles producing single peaks with a z-average of approximately 100 nm and below were used.

Birchenough H.L., Swann M.J., Zindy E., Day A.J, & Jowitt T.A. (2020). Enhanced avidin binding to lipid bilayers using PDP-PE lipids with PEG-biotin linkers. Nanoscale Advances, 2(4), 1625-1633.

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Endosomal Membrane Lipid Liposomes

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Phospholipids were selected based on phospholipids typically found in endosomal membrane except for charged phospholipids^{22 (link)}. POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine), POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine), cholesterol, and total gangliosides (GA) (Avanti Polar Lipids) were dissolved in chloroform-methanol (2:1) at a final concentration of 10 mM. Liposomes with 3 different lipid compositions were prepared: A) 95 mol % POPC and 5 mol % GA; B) 66 mol % POPC, 13 mol % POPE, 16 mol % cholesterol and 5 mol % GA; C) 42 mol % POPC, 13 mol % POPE, 40 mol % cholesterol and 5 mol % GA. The solvent was removed by evaporation; first using a stream of argon gas and second, high vacuum for 1 hour. The lipid film was re-suspended in KHE buffer, processed using 21 freeze-thaw cycles, and extruded at room temperature through 100 nm PC membrane (Whatman) using a mini-extruder set (Avanti Polar Lipids). Liposomes were stored at 4°C and used within 1 to 5 days.

Chlanda P., Mekhedov E., Waters H., Schwartz C.L., Fischer E.R., Ryham R.J., Cohen F.S., Blank P.S, & Zimmerberg J. (2016). The hemifusion structure induced by Influenza virus haemagglutinin is determined by physical properties of the target membranes. Nature microbiology, 1(6), 16050.

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Preparation of Stealth Copper-Loaded Liposomes

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Stealth liposomes were prepared by the hydration-extrusion method by incorporating 5% PEGylated lipids (DPPE-PEG₂₀₀₀) into the total phospholipid concentration (17 (link)). Briefly, lipid mixtures of DPPC, cholesterol and DPPE-PEG₂₀₀₀ at a molar ratio of 80:50:4 (µmol) were dissolved in chloroform and the solvent was evaporated on a rotary evaporator under vacuum. Non-PEGylated liposomes were prepared without the addition of DPPE-PEG₂₀₀₀. Next, 100 mM (or other concentrations) of CuCl₂ (or CuSO₄) solution was used to hydrate the thin film of lipids that formed on the wall of the round bottom flask, while rotating in a 55 °C water bath for 30 minutes. The resulting multilamellar vesicle (MLV) suspension was subjected to three freeze-thaw cycles before being extruded 11 times through a 100 nm polycarbonate membrane filter (Whatman, Springfield Mill, UK) via a Mini-Extruder Set (Avanti Polar Lipids) to generate large unilamellar vesicles (LUVs) encapsulating the Cu salt. Residual un-encapsulated Cu was removed by size exclusion chromatography by passing the suspension through a desalting PD-10 column (GE Healthcare, Waukesha, WI) and eluting with either ddH₂O (for short-term storage and in vitro experiments) or 0.9% sodium chloride (for in vivo administrations into mice). Saline liposomes were prepared by encapsulating normal saline solution instead of CuCl₂ solution.

Wang Y., Zeng S., Lin T.M., Krugner-Higby L., Lyman D., Steffen D, & Xiong M.P. (2014). Evaluating the Anticancer Properties of Liposomal Copper in a Nude Mouse Xenograft Model of Human Prostate Cancer: Formulation, In Vitro, In Vivo, Histology and Tissue Distribution Studies. Pharmaceutical research, 31(11), 3106-3119.

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Endosomal Membrane Lipid Liposomes

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