400 nm diameter polycarbonate filter

Manufactured by Avestin

The 400-nm-diameter polycarbonate filter is a laboratory equipment designed for filtration purposes. It is made of polycarbonate material and has a diameter of 400 nanometers. The core function of this filter is to separate and retain particles or substances based on their size.

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Lab products found in correlation

Poly prep column, by Bio-Rad (2 mentions)

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Polycarbonate filters (11 citations) 400 nm polycarbonate filter (3 citations)

2 protocols using 400 nm diameter polycarbonate filter

Reconstitution of Glutamate Transporter in Liposomes

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A solution of E. coli total lipid extract (20 mg ml⁻¹ in 50 mM KPi, pH 7.0) was extruded with a 400-nm-diameter polycarbonate filter (Avestin, 11 passages) and diluted with the same buffer to a final concentration of 4 mg ml⁻¹. The lipid mixture was destabilized with 10% Triton-X100. Purified Glt_Tk and the destabilized lipids were mixed in a ratio of 1:1600 or 1:250 (protein: lipid) and incubated at room temperature for 30 min. Bio-beads were added four times (25 mg ml⁻¹, 15 mg ml⁻¹, 19 mg ml⁻¹, 4 mg ml⁻¹ lipid solution) after 0.5 hr, 1 hr, overnight and 2 hr incubation, respectively, on a rocking platform at 4°C. The Bio-beads were removed by passage over an empty Poly-Prep column (Bio-Rad). The proteoliposomes were collected by centrifugation (20 min, 298,906 g, 4°C), subsequently resuspended in 50 mM KPi, pH 7.0 to the concentration of the protein 33.4 µg ml⁻¹ and freeze-thawed for four cycles. The proteoliposomes were stored in liquid nitrogen until subsequent experiments.

Arkhipova V., Trinco G., Ettema T.W., Jensen S., Slotboom D.J, & Guskov A. (2019). Binding and transport of D-aspartate by the glutamate transporter homolog GltTk. eLife, 8, e45286.

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Reconstitution of Glt Tk in Lipid Vesicles

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A solution of E. coli total lipid extract (20 mg ml⁻¹ in 50 mM KPi, pH 7.0) was extruded with a 400-nm-diameter polycarbonate filter (Avestin, 11 passages) and diluted with the same buffer to a final concentration of 5 mg ml⁻¹. The lipid mixture was destabilized with 10% Triton-X100. Purified Glt_Tk and the destabilized lipids were mixed in a ratio of 1:250 (protein: lipid) and incubated at room temperature for 30 min. Bio-beads were added four times (25 mg ml⁻¹, 15 mg ml⁻¹, 19 mg ml⁻¹, 29 mg ml⁻¹ lipid solution) after 0.5 hr, 1 hr, overnight and 2 hr incubation, respectively, on a rocking platform at 4°C. The Bio-beads were removed by passage over an empty Poly-Prep column (Bio-Rad). The proteoliposomes were collected by centrifugation (20 min, 298,906 g, 4°C), subsequently resuspended in 50 mM KPi, pH 7.0 to the lipid concentration of 20 mg ml⁻¹ and freeze-thawed for three cycles. The proteoliposomes were stored in liquid nitrogen until subsequent experiments.

Zhou W., Trinco G., Slotboom D.J., Forrest L.R, & Faraldo-Gómez J.D. (2021). On the Role of a Conserved Methionine in the Na+-Coupling Mechanism of a Neurotransmitter Transporter Homolog. Neurochemical Research, 47(1), 163-175.

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