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Teflon film

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Teflon film is a thin, durable, and chemically resistant material made from polytetrafluoroethylene (PTFE). It is known for its exceptional non-stick properties and is widely used in laboratory equipment and industrial applications.

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Lab products found in correlation

Hexadecane, by Merck Group (2 mentions) Chromar, by Mallinckrodt (2 mentions) Axopatch 200b, by Molecular Devices (2 mentions) Urea, by Thermo Fisher Scientific (1 mentions) Pentane, by Merck Group (1 mentions) 1 2 diphytanoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions) Clampfit 10, by Molecular Devices (1 mentions) Axopatch 200b amplifier, by Molecular Devices (1 mentions) Digidata 1440a converter, by Molecular Devices (1 mentions) Ni no3 2, by Merck Group (1 mentions) Ca no3 2, by Merck Group (1 mentions) Hg no3 2, by Merck Group (1 mentions) Th no3 4, by Merck Group (1 mentions) Zn no3 2, by Merck Group (1 mentions) Ascl3, by Merck Group (1 mentions) Mg no3 2, by Merck Group (1 mentions) Pb no3 2, by Merck Group (1 mentions) Hydrochloric acid (hcl), by Merck Group (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Trizma base, by Merck Group (1 mentions)

9 protocols using teflon film

1

Lipid Membrane Preparation and Characterization

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Lipids, 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC) and diphytanoylphosphatidylcholine (DPhPC) were purchased from Avanti Inc. Organic solvents (chloroform, spectral grade) and sodium chloride (Analytical grade) were purchased from Sigma Aldrich. Urea was obtained from Fisher Scientific. Carboxyfluoresceins were purchased from Sigma Aldrich. Teflon film was obtained from Goodfellow. Hexadecane and pentane were obtained from Sigma Aldrich.
Lin M., Zhang G., Fahie M., Morgan L.K., Chen M., Keiderling T.A., Kenney L.J, & Liang J. (2017). Engineering a Novel Porin OmpGF via Strand Replacement From Computational Analysis of Sequence Motif. Biochimica et biophysica acta, 1859(7), 1180-1189.
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2

Planar Lipid Membrane Electrical Recording

Cited 2 times
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Electrical
recording were performed using planar lipid membranes as described
before.35 (link),36 (link) Briefly, a 25-μm-thick Teflon film
(Goodfellow Corporation, PA, USA) containing an orifice of approximately
70 μm separates the cis and trans compartments. To form the membranes, 10 μL of 5% hexadecane
in pentane is added to the Teflon film and the pentane is allowed
to evaporate. The reservoirs are filled with buffer and 10 μL
of 10 mg/mL 1,2-diphytanoyl-sn-glycero-3-phosphocholine
(Avanti Polar Lipids) in pentane. Membranes were spontaneously formed
using the Montal–Mueller method.37 (link) Ag/AgCl electrodes are placed in each compartment, with the ground
electrode in the cis side. WT FraC oligomers are
added to the cis side of the chamber.23 (link),24 (link) Upon pore insertion, the pore is characterized by measuring traces
at different voltages and taking an IV curve. The
substrate was added to the cis side of the chamber
and measured at −90 mV.
Restrepo-Pérez L., Huang G., Bohländer P.R., Worp N., Eelkema R., Maglia G., Joo C, & Dekker C. (2019). Resolving Chemical Modifications to a Single Amino Acid within a Peptide Using a Biological Nanopore. ACS Nano, 13(12), 13668-13676.
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3

Single-Channel Recording of αHL Nanopore

Cited 1 time
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The procedure for single channel recordings have been described previously.27 (link) Briefly, a Teflon film (Goodfellow Malvern, PA) with a 150-μm diameter orifice separated two Teflon chamber compartments. Planar bilayer was formed according to the Montal-Muller method. Unless otherwise noted, the experiments were performed at 24 ± 1 °C using the wild-type αHL protein nanopore under symmetrical buffer conditions with the two chamber compartments filled with a solution consisting of 1 M NaCl, and 10 mM Tris (pH 7.5). Both the αHL protein and DNA polymers were added to the cis chamber compartment. The applied potential was +120 mV, unless otherwise noted. Ionic currents were recorded with Axopatch 200B amplifier (Molecular Devices, Sunnyvale, CA), filtered with a four-pole low-pass Bessel filter at 5 kHz, and then digitized with a Digidata 1440A converter (Molecular Devices) at a sampling frequency of 10 kHz. An average of 450 events was recorded in each of the single channel recording experiments. The event blockage amplitude, residence time, and number of occurrences (i.e., event counts) were obtained by using Clampfit 10.5 software (Molecular Devices).
Chen X., Roozbahani G.M., Ye Z., Zhang Y., Ma R., Xiang J, & Guan X. (2018). Label-free Detection of DNA Mutations by Nanopore Analysis. ACS applied materials & interfaces, 10(14), 11519-11528.
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4

Membrane Protein Reconstitution Protocol

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All chemicals were obtained from Fisher Scientific or Boston Bioproducts unless otherwise stated. Chemicals were used without further purification. SB58C mouse monoclonal antibody was obtained from Southern Biotech (Cat# 6406-01) and BTN.4 mouse monoclonal antibody was obtained from Thermo Scientific (Cat# MS-1048-P1). Diphytanoylphosphatidylcholine (DPhPC) lipids were obtained from Avanti Polar Lipids. Teflon film was obtained from Goodfellow. The maleimide-PEG2-biotin linker was obtained from Thermo Scientific. Octylglucoside (OG) was obtained from Gold Bio-technology. Hexadecane and pentane were obtained from Sigma Aldrich.
Fahie M.A., Yang B., Mullis M., Holden M.A, & Chen M. (2015). Selective detection of protein homologues in serum using an OmpG nanopore. Analytical chemistry, 87(21), 11143-11149.
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5

DNA Purification and Electrolyte Solutions

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DNA samples
with standard purification (desalting) were purchased from Intergrated
DNA Technologies (Coralville, IA). All the other chemicals were ordered
from Sigma-Aldrich (St. Louis, MO). All of the DNA samples and chemicals
were dissolved in HPLC-grade water (ChromAR, Mallinckrodt Baker).
All the stock solutions of DNA polymers were prepared at 5 mM each,
and kept at −20 °C before and after use. Three electrolyte
solutions were used in this work, which contained 0.15/1.0/3.0 M NaCl
buffered with 10 mM Trizma base, with the pH adjusted to 7.5 using
hydrochloride acid. Lipid 1, 2-diphytanoylphosphatidylcholine was
obtained from Avanti Polar Lipids (Alabaster, AL). Teflon film (25
μm thick) was purchased from Goodfellow (Malvern, PA).
Wang L., Han Y., Zhou S., Wang G, & Guan X. (2014). Nanopore Biosensor for Label-Free and Real-Time Detection of Anthrax Lethal Factor. ACS Applied Materials & Interfaces, 6(10), 7334-7339.
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6

Planar Lipid Membrane Nanopore Characterization

Cited 1 time
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Electrical recording was performed using planar lipid membranes (BLMs) as has been described before. 67 (link),68 (link) Briefly, a 25 um-thick Teflon film (Goodfellow Corporation, Pennsylvania USA) containing an orifice of approximately 70 um separates the cis and trans compartments. To form the membranes, 10 ul of 5% hexadecane in pentane is added to the Teflon film and the pentane is allowed to evaporate. The reservoirs are filled with buffer and 10 ul of 10 mg/ml DPhPC in pentane. Membranes were spontaneously formed using the Montal-Mueller method. Ag/AgCl electrodes are placed in each compartment, with the ground electrode in the cis side. WT FraC oligomers are added to the cis side of the chamber. Upon pore insertion, the pore is characterized by measuring traces at different voltages and taking an I-V curve. For the single-channel conductance measurements, nanopores were measured at 0, −50 and 50 mV. The substrate was added to the cis side of the chamber and measured at multiple voltages.
Zhao S., Restrepo-Pérez L., Soskine M., Maglia G., Joo C., Dekker C, & Aksimentiev A. (2019). Electro-Mechanical Conductance Modulation of a Nanopore Using a Removable Gate. ACS nano, 13(2), 2398-2409.
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7

Peptide D-12 Binding Interaction Assays

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Peptide D-12, a 12-amino acid peptide with a sequence of YEVHHQKDDPDD, was obtained from WatsonBio sciences (Houston, TX). Other chemicals such as Th(NO3)4 (99.999%), UO2(NO3)2 (99.999%), Ca(NO3)2 (99.999%), Mg(NO3)2 (99.999%), Ni(NO3)2 (99.999%), Zn(NO3)2 (99.999%), Cu(NO3)2 (99.999%), As(Cl)3 (99.999%), Pb(NO3)2 (99.999%), Hg(NO3)2(99.999%), NaCl (99.999%), HCl (ACS reagent, ≤1 ppm heavy metals), NaH2PO4 (BioXtra grade, ≥99.5%), and Trizma base (BioXtra grade, ≥99.9%) were bought from Sigma (St. Louis, MO). All the chemicals, including the D-12 peptide, were dissolved in HPLC-grade water (ChromAR, Mallinckrodt Baker). The stock solutions of the peptide and metal salts were prepared at concentrations of 10 mM each, and were kept at −20 °C before and after use. The buffer solutions used in this study included: (1) 1.0 M NaCl and 1 mM tris with pH values adjusted to 6.5 using HCl; (2) 1.0 M NaCl and 1 mM NaH2PO4 with pH values adjusted to 2.5, 3.5, 4.0, 4.5 and 5.5 using HCl. Lipid 1,2-diphytanoylphosphatidylcholine was purchased from Avanti Polar Lipids (Alabaster, AL). Teflon film was obtained from Goodfellow (Malvern, PA). The α-hemolysin (α-HL) (M113F)7 protein pores was made according to our previous work.35
Roozbahani G.M., Chen X., Zhang Y., Juarez O., Li D, & Guan X. (2018). Computation-assisted nanopore detection of thorium ions. Analytical chemistry, 90(9), 5938-5944.
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8

Single-Molecule Recordings in Lipid Bilayers

Cited 1 time
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We carried out single-molecule electrical recordings in planar lipid bilayers made of 1,2-diphytanoyl-sn-glycero-3-phophocholine (Avanti Polar Lipids) by the Montal and Mueller method across an aperture, 50-100 μm in diameter, in a 25-μm-thick Teflon film (Goodfellow) that had been painted with 1% hexadecane in pentane31 (link). The film separated two compartments (cis and trans), which were connected to the headstage of an amplifier Axopatch 200B (Molecular Devices) with Ag/AgCl electrodes, cis grounded. The amplifier was connected to a digitizer (Digidata 1440 A, Molecular Devices). A PC was used to collect data and control the amplifier with Clampex 10.5 software (Molecular Devices). Both the cis and trans compartments were filled with 2 M KCl, 10 mM Hepes, pH 7.2, at 22 ± 1.5 °C. When required, the cis compartment was supplemented with 5 mM nickel sulfate or 1 mM EDTA (final concentrations). After formation of a bilayer, 1 μL of heptamerized α-HL was added to the cis compartment. Following the first insertion of a pore, the remaining heptamerized α-HL was removed by perfusion with 10 chamber volumes of recording buffer. The signal was filtered with a low-pass Bessel filter of 5 kHz and data were collected at a sampling rate of 20 kHz.
Rosen C.B., Bayley H, & Rodriguez-Larrea D. (2020). Free-energy landscapes of membrane co-translocational protein unfolding. Communications Biology, 3, 160.
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9

Planar Lipid Bilayer Formation and Ionic Current Measurement

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DPhPC was used to form a stable lipid bilayer across an aperture 100 μm in diameter in a 25-μm-thick Teflon film (Goodfellow, Malvern, PA) that divided a planar bilayer chamber into two compartments, cis and trans. Both compartments contained 1 mL of KCl/CaCl2 buffer solution. Samples were added in the cis compartment, which was connected to the ground. The trans side was connected to the head-stage of the amplifier. Ionic currents were measured by using Ag/AgCl electrodes with a patch-clamp amplifier (Axopatch 200B; Axon instruments, Foster City, CA), filtered by a low-pass Bessel filter with a corner frequency of 5 kHz and then digitized with a Digidata 1440A A/D converter (Axon Instruments) at a sampling frequency of 100 kHz. The transmembrane potentials were mentioned in the article.
Peng W., Yan S., Zhou K., Wu H.C., Liu L, & Zhao Y. (2023). High-resolution discrimination of homologous and isomeric proteinogenic amino acids in nanopore sensors with ultrashort single-walled carbon nanotubes. Nature Communications, 14, 2662.
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