The amounts of cholesterol in whole cells and isolated plasma membranes were biochemically measured according to previously described methods (10 (link)). Plasma membranes were prepared as described previously (55 (link)). Briefly, cells were homogenized with a Teflon homogenizer (Wheaton) and centrifuged at 1,000 × g for 10 min. The postnuclear supernatant was collected, and the plasma membrane fraction was isolated using 30% Percoll-gradient fractionation (84,000 × g for 30 min) and extracted with chloroform/isopropyl alcohol/Nonidet P-40 (7:11:0.1). The apical and basal membranes were isolated from cultured HPAECs using cationic colloidal silica (Nalco 1060) and polyacrylic acid sodium salt (molecular weight ∼ 5,100; Sigma-Aldrich), according to a previously reported procedure (56 (link)). Free cholesterol and cholesteryl ester (esterified cholesterol) in the apical and basal plasma membrane fractions were assayed spectrophotometrically using a cholesterol quantitation kit (BioVision Inc.) and a microplate reader (ARVO SX; Perkin-Elmer Life Sciences) according to the manufacturers’ instructions and normalizing the value according to the protein concentration and cell number.
Polyacrylic acid sodium salt
Manufactured by Merck Group
Sourced in United States, Germany
Polyacrylic acid sodium salt is a chemical compound used in a variety of laboratory applications. It serves as a dispersant, emulsifier, and thickening agent. The compound is soluble in water and is commonly used in the formulation of various laboratory reagents and solutions.
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Lab products found in correlation
Ammonium hydroxide solution, by Merck Group (2 mentions)
Ac devd amc, by Enzo Life Sciences (2 mentions)
Cytotoxicity detection kit ldh release assay, by Roche (2 mentions)
Stabilized hydrogen peroxide solution 30 h2o2, by Merck Group (2 mentions)
Cm h2dcfda assay, by Thermo Fisher Scientific (2 mentions)
N acetylcysteine nac, by Merck Group (2 mentions)
6 hydroxydopamine hydrochloride, by Merck Group (2 mentions)
Ammonium cerium 4 nitrate, by Merck Group (2 mentions)
Cerium 3 nitrate hexahydrate, by Merck Group (2 mentions)
Propidium iodide, by Merck Group (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Fluorobrite dmem, by Thermo Fisher Scientific (2 mentions)
Cholesterol quantitation kit, by Abcam (1 mentions)
Arvo sx, by PerkinElmer (1 mentions)
Ludox hs 40, by Merck Group (1 mentions)
Sucrose c12h22o11, by Merck Group (1 mentions)
Carbon black n550, by Cabot (1 mentions)
Calcium acetate monohydrate ca oac 2 h2o, by Merck Group (1 mentions)
Milli q plus 185, by Merck Group (1 mentions)
Aphidicolin, by Merck Group (1 mentions)
11 protocols using polyacrylic acid sodium salt
1
Cholesterol Measurement in Plasma Membranes
2
Synthesis and Characterization of Silica and Polyacrylic Acid Nanoparticles
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Silica NPs were acquired from Nanotesla Institute Ljubljana (Ljubljana, Slovenia). Silica NPs were dispersed in a solution of citric acid in ethanol at pH 2. Z-average size of silica NPs in dispersion was 58.6 nm with polydispersity index (PDI) 0.46. PAA NPs were prepared as described elsewhere [38 (link)]. Briefly, cobalt ferrite (Co ferrite) NPs were prepared by co-precipitation [39 ] and stabilized in water. Alkaline medium was removed and NPs were re-suspended in distilled water. The process was repeated three times and after the last washing step nitric acid was added to stabilize NPs in acidic media. NPs were coated in situ with a 45% (m/m) water solution of polyacrylic acid sodium salt with molecular mass of 8 kDa (Sigma-Aldrich, St. Luis, Missouri, USA), by mixing 10 ml of ferro fluid and 10 ml of PAA water solution of equal mass concentrations at 20 mg/ml for 10 min at room temperature. Unbound polyacrylic acid was removed with dialysis against dH2O. dH2O was replaced every four hours, four times in total, to ensure removal of unbound polyacrylic acid. Larger aggregates were removed by filtration with Minisart® filter unit (Sartorius Stedim Biotech, Göttingen, Germany), the size of pores was 0.20 μm. Z-average size of PAA NPs in dispersion was 116.2 nm with PDI 0.18.
3
Preparation of Reinforced EPDM Composites
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Ludox HS-40 (40 wt%, 12 nm), sucrose (C12H22O11, 99.5%), and poly (acrylic acid sodium salt) were provided by Sigma Aldrich (St. Louis, MO, USA). EPDM was obtained from the Jilin Chemical Industrial Limited Company of China (Jilin, China). Carbon black N550 was obtained from Cabot Corporation (St. Boston, MA, USA) in the United States; foaming agent AC was acquired from Changzhou Yongxin Fine Chemical Co., Ltd. (Changzhou, China). The antioxidant 2246, zinc oxide (ZnO), stearic acid (SA), accelerator N-cyclo-hexylbenzothiazole-2-sulphenamide (CZ), sulfuric acid (H2SO4, 98%), sodium hydroxide (NaOH), and insoluble sulfur (S) are all industrial grade products from Kemiou Chemical Co., Ltd. (Tianjin, China).
4
Synthesis of Calcium Acetate Hydrogels
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All chemical reagents are commercially available. Calcium acetate monohydrate (Ca(OAc)2H2O) and poly(acrylic) acid sodium salt (average MW = 8000 Daltons) were purchased from Sigma Aldrich (USA). The water used in all experiments was prepared in a three-stage Millipore MilliQ plus 185 purification system, and had a resistivity of 18.2 mΩ cm.
5
Antibodies for DNA Replication Proteins
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A commercial antibody was used to detect ORC1 in extracts: anti-ORC1 (clone TK1/2, cat.# sc-53391, Santa Cruz). Antibodies against ORC2, CDC6, MCM and RPA were generated against the corresponding polypeptides and previously described (51 (link),52 (link)). Mouse anti-phospho-Histone H2AX (Ser139, JBW301 #05–636; Millipore), rabbit anti-histone H3 (#9715; Cell Signaling), rabbit anti–CHK1-pS345 (#2341; Cell Signaling) and mouse anti-phospho-ATM (Ser1981, #200-301-400 Rockland) were purchased from commercial sources.
Silver nitrate (AgNO3, 99.9995%) and sodium borohydride (NaBH4, 98%) were purchased from Alfa Aesar. Poly(acrylic acid) sodium salt 35 wt% (MW≈15 000) and 3-aminopropyltriethoxysilane (APTES) were obtained from Sigma-Aldrich. α-[32P]dCTP was purchased from Amersham Biosciences. ANTI-FLAG® M2 Affinity Agarose Gel and aphidicolin were purchased from Sigma-Aldrich. SYBR–Gold was purchased from Molecular Probes. All reagents were of the analytical reagent grades and utilized as received. All samples were prepared using ultrapure water (18.2 MΩ; Millipore Co.).
Silver nitrate (AgNO3, 99.9995%) and sodium borohydride (NaBH4, 98%) were purchased from Alfa Aesar. Poly(acrylic acid) sodium salt 35 wt% (MW≈15 000) and 3-aminopropyltriethoxysilane (APTES) were obtained from Sigma-Aldrich. α-[32P]dCTP was purchased from Amersham Biosciences. ANTI-FLAG® M2 Affinity Agarose Gel and aphidicolin were purchased from Sigma-Aldrich. SYBR–Gold was purchased from Molecular Probes. All reagents were of the analytical reagent grades and utilized as received. All samples were prepared using ultrapure water (18.2 MΩ; Millipore Co.).
6
Cerium Nanoparticle Synthesis and Characterization
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Cerium (III) nitrate hexahydrate, ammonium cerium (IV) nitrate, polyacrylic acid sodium salt (Mw = 5100), ammonium hydroxide solution (30% NH3 in H2O) and fluorescein 5-isothiocyanate were purchased from Sigma-Aldrich (Germany). For cell culture, Dulbecco's Modified Eagle Medium (DMEM), Trypsin/EDTA (0.25%) solution, heat-inactivated fetal bovine serum (FBS), Dulbecco's phosphate buffered saline (DPBS, without calcium and magnesium) and FluoroBrite™ DMEM were procured from Gibco (Invitrogen, Paisley, UK). The cytotoxicity detection kit (LDH release assay) was from Roche Diagnostic (Switzerland). Penicillin and streptomycin mixture, Triton X-100, N-acetyl cysteine (NAC), Ac-DEVD-CHO, propidium iodide, stabilized hydrogen peroxide solution (30% H2O2) and 6-hydroxydopamine hydrochloride were from Sigma Aldrich (Germany). Ac-DEVD-AMC, a caspase-3 fluorogenic substrate was obtained from Enzo Life Sciences (New York, NY, USA). CM-H2DCFDA assay was purchased from Molecular Probes (USA).
7
Upconversion Particle Glucose Sensing
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Example 1
Commercially available UCP particles (Honeywell Lumilux Green UC 2, Honeywell Specialty Chemicals Seelze GmbH) were mixed with polyacrylic acid (poly(acrylic acid, sodium salt), Sigma-Aldrich) and then embedded in to the bottom chemical layer by mixing them with the standard coating mixture as described below. These particles were composed of Y2O2S:Yb,Er and were excited with an infrared radiation (980 nm). The test strips were then used to investigate the correlation between the detected UCP emission as a function of glucose concentration under infrared laser excitation.
8
Synthesis of Thermosensitive PNIPAM/PAA Microgels
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The PNIPAM/PAA microgel was formed by surfactant-free precipitation polymerization. A monomer of NIPAM (9 × 10−2 mol, N-Isopropylacrylamide > 98%, TCI, Chennai, India) was adequately purified and then recrystallized using hexane, polyacrylic acid sodium salt (1.6 × 10−2 mol, MW 15,000 g/mol, Sigma-Aldrich, Darmstadt, Germany) and MBA (2.6 × 10−4 mol N,N-Methylenebisacrylamide > 99%, Alfa Aesar, Ward Hill, MA, USA), which was dissolved in an aqueous solution (final volume at 200 mL). Acting as a cross-linker for this synthesis, MBA was selected. The reaction mixture was then purged for 1 h with pure N2 (99.99999%) and afterward heated to 70 °C. After this step, KPS (7 × 10−5 mol potassium persulfate > 99%, Fisher Chemicals, Hampton, NY, USA) was also added and the polymerization process, starting at a duration of 6 h, under vigorous stirring (Figure 1 ). The size behavior of the microgels was estimated by dynamic light scattering (DLS) at 173° optics (Zeta Sizer nano S, Malvern Inst., Malvern, UK).
9
Synthesis and Cytotoxicity Evaluation of Rare-Earth Nitrates
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Cerium (III) nitrate hexahydrate, ammonium cerium (IV) nitrate, europium (III) nitrate hydrate, polyacrylic acid sodium salt (Mw = 5100) and ammonium hydroxide solution (30% NH3 in H2O) were procured from Sigma-Aldrich (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany). For cell culture, Dulbecco’s Modified Eagle Medium (DMEM), Trypsin/EDTA (0.25%) solution, heat-inactivated fetal bovine serum (FBS), Dulbecco’s phosphate-buffered saline (DPBS, without calcium and magnesium), penicillin and streptomycin mixture and FluoroBrite™ DMEM were procured from Gibco (Invitrogen, Paisley, UK). The cytotoxicity detection kit (LDH release assay) was from Roche Diagnostic (Mannheim, Germany). Ac-DEVD-AMC, a caspase-3 fluorogenic substrate, was obtained from Enzo Life Sciences (New York, NY, USA). CM-H2DCFDA assay was purchased from Molecular Probes (Life Technologies Corporation, Eugene, OR, USA). Triton X-100, N-acetyl cysteine (NAC), propidium iodide, stabilized hydrogen peroxide solution (30% H2O2) and 6-hydroxydopamine hydrochloride were sourced from Sigma Aldrich (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany).
10
Upconverting Particle-based Glucose Sensing
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Example 1
Commercially available UCP particles (Honeywell Lumilux Green UC 2, Honeywell Specialty Chemicals Seelze GmbH) were mixed with polyacrylic acid (poly(acrylic acid, sodium salt), Sigma-Aldrich) and then embedded in to the bottom chemical layer by mixing them with the standard coating mixture as described below. These particles were composed of Y2O2S:Yb,Er and were excited with an infrared radiation (980 nm). The test strips were then used to investigate the correlation between the detected UCP emission as a function of glucose concentration under infrared laser excitation.
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