All the chemicals were used as procured unless specified. Polycaprolactone (Mw = 70 000 GPC; Scientific Polymer Products, USA), PVDF (Mw = 534 000; Sigma Aldrich; France) and PANI (emeraldine base; avg Mw ∼ 50 000; Aldrich Chemistry, USA) were used for synthesis of the scaffolds including PCL and PCL–PVDF. The doped form of PANI–PCL solution was prepared by dissolving equal mass of emeraldine base and 10-camphorsulfonic acid (HCSA) (Aldrich Chemistry; France) in a solution of 10% PCL in chloroform. The PCL–PANI solution was filtered to remove suspensions larger than 0.22 μm since the doped form of PANI is immiscible.21 DMF (Macron Fine Chemicals; USA) and acetone (Sigma Aldrich, USA) in the ratio 7 : 3 were used to prepare a 20% PVDF solution. PCL–PANI–PVDF scaffold was prepared by coaxially electrospinning from separate syringes which were controlled using syringe pumps (Harvard Pumps, EP-H11). The PCL–PANI (core solution) and 20% PVDF (sheath solution) were electrospun (EM-DIG; IME Technologies, Netherlands) at 18 kV using a rotating collector at a rotational speed of 750 rpm. The parameters used for fabrication of PCL and PCL–PVDF scaffolds are elaborated in the ESI.†
Polyvinylidene fluoride (pvdf)
Manufactured by Merck Group
Sourced in United States, Germany, China, United Kingdom, India, Ireland, Morocco, Italy, Japan, Macao, France, Canada
PVDF is a type of laboratory equipment used for various applications. It is a fluoropolymer material with a unique set of properties, including chemical resistance, thermal stability, and mechanical strength. PVDF is commonly used in the manufacturing of laboratory equipment, such as filter membranes, tubing, and other components that require these specific characteristics.
Automatically generated - may contain errors
Lab products found in correlation
Ripa lysis buffer, by Beyotime (76 mentions)
Bca protein assay kit, by Beyotime (71 mentions)
Ripa buffer, by Beyotime (70 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (66 mentions)
Ripa buffer, by Thermo Fisher Scientific (34 mentions)
β actin, by Cell Signaling Technology (31 mentions)
Protease inhibitor cocktail, by Roche (31 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (31 mentions)
Image lab software, by Bio-Rad (29 mentions)
Radioimmunoprecipitation assay (ripa), by Beyotime (28 mentions)
N methyl 2 pyrrolidone, by Merck Group (28 mentions)
β actin, by Santa Cruz Biotechnology (26 mentions)
Quantity one software, by Bio-Rad (26 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (25 mentions)
Gapdh, by Abcam (24 mentions)
Bca kit, by Thermo Fisher Scientific (24 mentions)
β actin, by Abcam (22 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (22 mentions)
Pvdf membrane, by Merck Group (21 mentions)
Ripa buffer, by Merck Group (21 mentions)
1 776 protocols using polyvinylidene fluoride (pvdf)
1
Fabrication of PCL-PANI-PVDF Scaffolds
2
Nile Red-Doped PVDF Thin Films
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Thin films of nile red (NR, Aldrich,
99+% pure by LC-MS) embedded in poly(vinylidene fluoride) (PVDF, Sigma-Aldrich,
MW ≈ 534 000 by GPC) were prepared as described previously.18 (link) Films were ∼300 nm thick as determined
by ellipsometry, with sample preparation tailored to express the ferroelectric
β phase of PVDF, as described in the literature.19 Heavily dyed samples demonstrated no degradation
in fluorescence intensity or optical density over periods of months.
99+% pure by LC-MS) embedded in poly(vinylidene fluoride) (PVDF, Sigma-Aldrich,
MW ≈ 534 000 by GPC) were prepared as described previously.18 (link) Films were ∼300 nm thick as determined
by ellipsometry, with sample preparation tailored to express the ferroelectric
β phase of PVDF, as described in the literature.19 Heavily dyed samples demonstrated no degradation
in fluorescence intensity or optical density over periods of months.
3
Fabrication of KNTO@Ag-PVDF Composites
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To prepare three-phase KNTO@Ag-PVDF polymer composites, a 2 wt.% solution of poly (vinylidene fluoride) (PVDF, Sigma Aldrich, Mw~530,000, St. Louis, MO, USA) was first prepared in dimethylsulfoxide ((CH3)2SO, 99.5%, VitaHim, Dzerzhinsk, Russia), to which the corresponding KNTO@Ag powders were added in amounts of 7.5, 15, 22.5, and 30 vol.%. The theoretical density value of 3.878 g/cm3, calculated using the Rietveld method, was used to calculate the mass weights of KNTO powder. For homogenization, the resulting dispersions were subjected to mechanical stirring on a magnetic stirrer for 24 h, followed by ultrasonic treatment for 1 h. The prepared mixtures were poured into deionized water and dried at 100 °C for 10 h to remove the solvent. From the obtained composites, disks 12 mm in diameter and 1 mm in thickness were formed using uniaxial hot pressing at a temperature of 200 °C and a pressure of 20 MPa. Conductive silver paste was applied to the obtained composite products as electrodes.
4
Western Blot Analysis of Apoptosis Markers
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To test protein levels of p-ERK/p-JNK/p-P38 MAPK, Bax/Bcl-2, caspase-1, COX2, and iNOS in different groups, astrocyte lysates were homogenized by RIPA lysis buffer. Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene fluoride (PVDF, Millipore, Bedford, MA) membranes. The PVDF membranes were first blocked with 5% bull serum albumin (BSA) for 1 h at room temperature, then incubated with the primary antibody [dilution for GAPDH was 1: 8000, Sigma, St. Louis, MO; dilutions for Bax/Bcl-2, caspase-1, COX2, iNOS, JNK, ERK 1/2 and p-p38 (Tyr182) were 1: 1000, Cell Signalling Technology, Beverly, MA] at 4°C overnight. On the next day, the PVDF membranes were washed 3 times for 10 min each time with Tris-buffered saline Tween (TBST), then incubated with horseradish peroxidase (HRP)-conjugated secondary antibody for 1 h at room temperature. After washing 3 times with TBST, bands were developed with an ECL luminescence reagent. Glyceraldehyde-phosphate dehydrogenase (GAPDH) was the loading control.
5
Electrochemical Characterization of AT Anode
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The electrodes for the electrochemical measurements were fabricated by mixing the active material 1,4,9,10-anthracenetetraone (AT) with super P and polyvinylidene difluoride (PVDF, average M 275 000 g mol−1, Sigma-Aldrich) with a mass ratio of 8 : 1 : 1. In particular, the organic active material was ground with super P in an agate mortar for 15 minutes to obtain the black mixed powder, which was subsequently dispersed in N-methylpyrrolidone (NMP, ≤99%, Sigma-Aldrich) solution with the dissolved PVDF (0.05 g ml−1). The mixed slurry was treated by ultrasonication for 30 minutes and cast on Al foil. The half-cell consisted of a working electrode, lithium metal (99.9%, Sigma-Aldrich), and a glass fiber separator (ECC1-01-0012-B/L, EL-CELL), while the cell assembly was performed in an argon-filled glove box. The electrolyte is composed of a solution mixture of EC and DMC (v/v = 1 : 1) with 1.0 M LiPF6 (Sigma-Aldrich, battery grade). The cyclic voltammetry (CV) measurements were conducted using a galvanostat/potentiostat (VMP-300, BioLogic) with a scan rate of 0.2 mV s−1.
6
PVDF Solution Preparation and CNT Yarn
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The 24 wt % PVDF solution was prepared
using PVDF (Mw = 275 000 g mol–1, Sigma-Aldrich)
pellets dissolved in a 1:1 solution of dimethylacetamide (DMAc, analytical
standard, Avantor) and acetone (analytical standard, Avantor). The
solution was stirred for 4 h at a constant speed of 700 rpm on a hot
plate set to 50 °C (IKA RCT basic). The CNT yarn was purchased
from DexMat Inc. and used as received.
using PVDF (Mw = 275 000 g mol–1, Sigma-Aldrich)
pellets dissolved in a 1:1 solution of dimethylacetamide (DMAc, analytical
standard, Avantor) and acetone (analytical standard, Avantor). The
solution was stirred for 4 h at a constant speed of 700 rpm on a hot
plate set to 50 °C (IKA RCT basic). The CNT yarn was purchased
from DexMat Inc. and used as received.
7
Western Blotting for Alpha-Toxin and PVL
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After 12% SDS-PAGE separation of the samples, the proteins were transferred to PVDF (Millipore). PVDF membranes were then incubated with blocking buffer for 2 h. The rabbit anti-alpha-toxin polyclonal antibody (diluted 1:10,000) (Cat number S7531, Sigma-Aldrich) and the rabbit polyclonal anti-PVL LukS subunit (0.5 μg/mL, Cat number ab190473, Abcam, Cambridge, United Kingdom) were used to detect alpha-toxin and PVL, respectively. The HRP-labeled goat anti-rabbit IgG (diluted 1:2,000) (Cat number SE134, Solarbio, Beijing, China) was used as the secondary antibody. Western blotting bands were quantified using Image J Software.
8
Mitochondria Fraction Purity Validation
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The integrity and purity of the mitochondria fraction were confirmed by western blot analysis for several protein markers. Twenty micrograms of protein were separated by 4–12% gradient SDS-PAGE, transferred onto a PVDF membrane, electrotransferred to polyvinylidene difluoride membrane (PVDF) (Millipore, Billerica, MA), and probed with primary antibody followed by incubation with horseradish peroxidase-labeled IgG (1:5000) depending on the primary antibody. The purity of the mitochondria fraction was tested by screening for mitochondria protein utilizing the following antibodies: anti-VDAC (1:1000, 48661, cell signaling, CST), anti-β -tubulin (1:5000, ab21058, Abcam) for the cytosol, anti-calnexin for the nuclear and endoplasmic reticulum (2433 S, cell signaling, CST) in the isolated mitochondria versus in the supernatant fraction. We found that all mitochondria preparations were essentially free of cytosolic and nuclear contaminants. Together, these results attest to the purity of our mitochondria fractions, i.e., they were substantially free of cytosolic, nuclear or endoplasmic reticulum components.
9
Western Blot Analysis of FLAG-tagged Proteins
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The cells were harvested and then lysed on ice for 20 minutes with a pre-cooled lysis buffer (RIPA Lysis Buffer III, Sangon Biotech, Shanghai, China). The lysate was centrifuged at 12,000 rpm at 4 °C for 15 minutes, and then the supernatant was collected. Protein concentration was measured using a BCA Protein Assay Kit (Beyotime, Shanghai, China). Ten percent SDS-PAGE gels were utilized to resolve 30 µg of protein from each sample. The proteins were then transferred onto polyvinylidene difluoride (PVDF) (Millipore, MA, USA) membranes. The membranes were blocked for 1 h with PBST containing 5% non-fat dry milk powder. The PVDF membranes were incubated with rabbit anti-FLAG primary antibody (1:1,000, SAB, Shanghai, China) overnight at 4 °C and then with goat anti-rabbit IgG (H+L) secondary antibody (1:1,000, Proteintech Group, Chicago, USA) for 2 hours at room temperature. The PVDF membranes were soaked in PBST buffer and washed slowly for 10 minutes. The PVDF membranes were placed on the plastic wrap, and then the ECL reagent (Millipore, MA, USA) was added to the PVDF membranes. The enhanced chemiluminescence assay was performed using the ChemiDoc XRS+ (Bio-Rad, Minnesota, USA) according to the manufacturer’s instructions. β-actin was used as a normalization control. Finally, the grayscale value analysis was carried out with ImageJ software.
10
Quantitative Protein Analysis via Western Blot
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Equal amounts of denatured proteins are extracted from cells or tissue homogenates with lysates containing protease inhibitors. Protein concentration is measured with the bicinchoninic acid assay (BCA) assay kit. Proteins were electrophoresed on polyacrylamide gels and transferred to polyvinylidene difluoride membranes (PVDF, Millipore). Proteins were electrophoresed on polyacrylamide gels and transferred to polyvinylidene difluoride membranes (PVDF, Millipore). PVDF membranes were blocked for 2 h at room temperature using tris buffered saline with tween 20 (TBST) containing 5% skim milk. Incubate overnight at 4°C with the addition of primary antibodies. Secondary antibodies are then added and incubated for 2 h at 37°C. Proteins on PVDF membranes were quantified using an enhanced chemiluminescence system (ECL) and subsequently analyzed using Image Quant TL software (GE).
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