A frustule suspension was used as filler. The gels were prepared with 10% and 20% of polyacrylamide using 30% acrylamide/N,N'-methylene-bis acrylamide solution (29:1) (Sigma-Aldrich, Milan, Italy), 3μl of 10% ammonium persulfate (APS) (BioRad) solution, with or without 1% w/v of frustules, 2.5μl of 0.1mg/ml Rho B solution (Sigma–Aldrich, Milan, Italy) and ddH2O for a final volume of 65μl. The polymerization was performed using 1 μl of tetramethylethylenediamine (TEMED) (BioRad) in O-ring of 5 mm diameter and 1mm of thickness in order to obtain gels with the same dimensions.
Ammonium persulfate aps
Manufactured by Bio-Rad
Sourced in United States
Ammonium persulfate (APS) is an oxidizing agent commonly used as an initiator in various chemical reactions, particularly in the polymerization process. It is a colorless, crystalline solid that is soluble in water. APS is a versatile chemical compound with a wide range of applications in the field of chemistry and materials science.
Automatically generated - may contain errors
Lab products found in correlation
Temed, by Bio-Rad (3 mentions)
Acrylamide, by Bio-Rad (2 mentions)
N n n n tetramethylethylenediamine temed, by Bio-Rad (2 mentions)
Tween 20, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Acrylamide bisacrylamide, by Bio-Rad (2 mentions)
Sodium periodate naio4, by Thermo Fisher Scientific (1 mentions)
N n methylenebis acrylamide mba, by Merck Group (1 mentions)
Fibronectin solution, by Merck Group (1 mentions)
Bovine plasma, by Merck Group (1 mentions)
Sulfo sanpah, by Merck Group (1 mentions)
Milli q ultrapure system, by Merck Group (1 mentions)
N n methylenebisacrylamide bis, by Merck Group (1 mentions)
Poly ethylene glycol dimethacrylate pegdma, by Merck Group (1 mentions)
Resolving gel buffer, by Bio-Rad (1 mentions)
Stacking gel buffer, by Bio-Rad (1 mentions)
Allprep dna rna protein kit, by Qiagen (1 mentions)
Anti tet3, by Abcam (1 mentions)
Acrylamide bis acrylamide solution, by Bio-Rad (1 mentions)
Anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions)
14 protocols using ammonium persulfate aps
1
Polyacrylamide Gels with Frustule Fillers
2
Hydrogel Synthesis Reagents Protocol
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N-isopropylacrylamide (NIPAM) (purity 98%) monomer was purchased from TCI (Tokyo, Japan). Acrylamide (AM) (purity > 99.5%) monomer was purchased from BioShop (Burlington, ON, Canada). Ammonium persulfate (APS) with a purity over 99% and N, N, N’, N’-tetramethylenediamine (TEMED) with a purity higher than 98% were purchased from Bio-Rad (Hercules, CA, USA). N, N’-methylenebis(acrylamide) (MBA) was obtained from Sigma Aldrich (St. Louis, MO, USA). Sodium periodate (NaIO4) (purity 98%) was purchased from Alfa Aesar (Ward Hill, MA, USA).
3
Polyacrylamide Gel Fabrication for Cell Culture
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Polyacrylamide (PA) gels were prepared on aminosilanized glass cover slips as previously described [14] (link). Briefly, 40% w/v acrylamide and 2% w/v bis-acrylamide stock solutions (Bio-Rad, Hercules, CA) were mixed to prepare a PA solution and then the gel’s stiffness was achieved by varying the final concentration of the PA solution (3% ∼7.5%) and Bis-acrylamide cross-linker (0.06% ∼0.4%). To polymerize the solutions, 2.5 µl of 10% w/v ammonium persulfate (APS; Bio-Rad, Hercules, CA) and 0.25 µl of N,N,N′,N′-Tetramethylethylenediamine (TEMED; Bio-Rad, Hercules, CA) were added to yield a final volume of 500 µl PA solution. To crosslink extracellular matrix molecules onto the gel surface, a photoactivating cross-linker, 0.5 mg/ml of sulfo-SANPAH (sulfosuccinimidyl 6-(4′-azide-2′-nitrophenyl-amino) hexanoate, Pierce, Rockford, IL) solution was used. The powder of sulfo-SANPAH was dissolved in 10 mM HEPES buffer containing 0.5% DMSO and the solution was added on top of the PA gel. Gel dishes were placed at a distance of ∼15 cm from the UV light of the hood for 6 min and rinsed three times with 10 mM HEPES buffer for 10 min. A 200 µl of fibronectin solution (5, 10 or 40 µg/ml) from bovine plasma (Sigma, St. Louis, MO) was incubated on top of the sulfo-SANPAH-coated PA gel at 37°C overnight to deposit fibronectin for subsequent cell seeding.
4
Hydrogel Synthesis Using Monomers and Crosslinkers
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The monomers polyethylene glycol methyl ether methacrylate (PEGMEM), Mw: 300, 500, and 950 g·mol−1 and 2-dimethylamino ethyl methacrylate (DMAEM), the crosslinkers N,N′-methylenebis(acrylamide) (BIS) and polyethylene glycol dimethacrylate (PEGDMA), Mw: 550, and the catalyst N,N,N′,N′-Tetramethylethylenediamine (TMED) were purchased from Sigma Aldrich, Darmstadt, Germany. The initiator ammonium persulfate (APS) was purchased from Bio-Rad, Tokyo, Japan. All chemicals were used without further purification. The water used in the preparation and dialysis was purified on a Milli-Q ultrapure system (Millipore, Molsheim, France).
5
Western Blot Analysis of TET1 and TET3
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Protein lysates were prepared using AllPrep DNA/RNA/Protein Kit (Qiagen), according to the manufacturer’s instructions. Polyacrylamide gels were poured in 8 % concentration using 30 % acrylamide/bisacrylamide solution (Bio-Rad), sodium dodecyl sulfate (SDS) (Bio-Rad), ammonium persulfate (APS) (Bio-Rad), tetramethylethylenediamine (TEMED) (Bio-Rad), Resolving Gel Buffer (Bio-Rad) and Stacking Gel Buffer (Bio-Rad). Electrophoresis of protein lysates was performed using 10 µL of each sample, followed by Ponceau S staining. Samples were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore). The following antibodies were used for the detection of TET1, TET3, and beta-actin: anti-TET1 in dilution of 1:1000 (Thermo Fisher, PA5-72805), anti-TET3 in dilution 1:1000 (Abcam, ab 139311) and anti-beta-actin in dilution of 1:10 000 (Cell Signaling, 4967L). All membranes were incubated with a secondary anti-rabbit IgG antibody in a dilution of 1:7500, conjugated to horseradish peroxidise (HRP) (Thermo Fisher). Protein signals were detected by chemiluminescence using Clarity Max Western ECL Substrate (Bio-Rad) and quantified with ImageLab software (Bio-Rad). TET1 and beta-actin measurements were normalized to whole-cell lysate content based on the Ponceau S staining.
6
Synthesis and Characterization of Gold Nanomaterials
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All the chemicals are commercially available and used without further purification. Gold(III) chloride trihydrate (HAuCl4·3H2O, 99.999%), L-Glutathione reduced (GSH) ≥ 98.0%, sodium cyanoborohydride (NaBH3CN, 95%), sodium borohydride (NaBH4, 99.99%), and aqueous suspension of gold nanorods (10 nm diameter, 780 nm SPR absorption maximum) were purchased from Sigma-Aldrich. Aqueous 40% acrylamide and bisacrylamide stock solution (37.5:1), N,N,N′,N′-tetramethylethylenediamine (TEMED, 99%), N,N′-methylene bisacrylamide (Bis, >98%), and 10× concentrated SDS-PAGE running buffer (SDS) were supplied from Carl Roth. Ammonium persulfate (APS, 98%), 0.5 M Tris-HCl buffer pH 6.8, and 1.5 M Tris-HCl buffer pH 8.8 were purchased from BIO-RAD. Deionized (milli-Q) water with a resistivity of 18 MΩ cm was used in this work.
7
Histone H1 Modification and Detection
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Histone H1, 2,4-dinitrophenyl hydrazine (DNPH), 1-anilinonaphthalene-8-sulfonic acid (ANS), sodium dodecyl sulfate (SDS), methylglyoxal, aminoguanidine hydrochloride, diethylene triamine penta-acetic acid (DTPA), sodium azide, ethidium bromide, protein A-agarose (2.5ml pre-packed column), agarose, sodium azide, Tween-20, dialysis tubings, anti-human and anti-rabbit IgG, alkaline phosphatase conjugate, para-nitrophenyl phosphate, Freund’s complete and incomplete adjuvants were purchased from Sigma Chemical Company, St. Louis, MO, USA. Acrylamide, bisAcrylamide, ammonium persulfate (APS) and N,N,N′,N′- tetramethylethylenediamine (TEMED) were from Bio-Rad Laboratories, U.S.A. Sodium hydroxide, Ethylenediaminetetraacetic acid (disodium salt), methanol, glacial acetic acid, iso-propanol, sodium chloride, sodium carbonate, sodium nitrite, silver nitrate, xylene, sodium hydroxide, formaldehyde, sodium bicarbonate, ethanol, ammonium sulphate and ammonium persulphate were obtained from Qualigens, India. Polystyrene microtitre flat bottom ELISA plates and modules were purchased from NUNC, Denmark. All other chemicals/reagents were of the highest analytical grade available.
8
Lactoferrin Conjugation and Characterization
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Glycyrrhizin (ammonium salt, purity 95%), sodium periodate (NaIO4), lactoferrin human (Lf), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), sodium cyanoborohydride (NaBH3CN), fluorescein isothiocyanate isomer 1 (FITC), 2-mercaptoethanol and tetramethylethylenediamine (TEMED), and water (HPLC grade) wer.e purchased from Sigma-Aldrich (St. Louis, MO, USA). 4′,6-diamidino-2-phenylindole (DAPI) was purchased from Vector Laboratories (Burlingame, CA, USA). NP40, cell extraction buffer, and Lysotracker® were purchased from Invitrogen (Waltham, MA, USA). Phenylmethylsulfonyl fluoride (PMSF), acetonitrile, and methanol (HPLC grade) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Paraformaldehyde (4%) was purchased by Wako (Kanto, Saitama, Japan). Protease inhibitor and Calcein AM (Cat #: BMD00064) were purchased from Abbkine (Wuhan, China). Sodium dodecyl sulfate was purchased from Affymetrix (Waltham, MA, USA). Acrylamide solution (30%) and ammonium persulfate (APS) were purchased by BIO-RAD (Hercules, CA, USA). Coomassie blue solution was purchased from Abcam (Cambridge, UK).
9
Antioxidant Activity of F. philippinensis
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Root barks of F. philippinensis were gathered from a farm in Ninning, Guanxi province, China. The plant was confirmed with Voucher specimens (No. 530) by Dr. Yimmin Zhao, Gaunxi Botanical Garden, China. It was cleaned, dried, and stored at a cool temperature (<18 °C) in darkness. Methanol, chloroform, butanol, n-hexane, sodium hydroxide (NaOH), and copper sulfate (CuSO4) were purchased from Duksan Co. (Gyeonggi-do, South Korea). xanthine oxidase from bovine milk (EC 1.17.3.2), xanthine, allopurinol, sodium pyrophosphate, dimethyl sulfoxide (DMSO), probucol, thiobarbituric acid, trichloroacetic acid, and other used organic solvent were from Sigma Aldrich Co., (St. Louis, MO, USA). Low-density lipoprotein (LDL) was taken from Invitrogen (Thermo Fisher Scientific, Carlsbad, California, USA). ODS-C18 and Triart prep C18-S were from YMC Co., (Kyoto, Japan). Agarose, sodium dodecyl sulfate (SDS), N,N,N′,N′-tetramethylethylenediamine (TEMED), coomassie brilliant blue R-250, ammonium persulfate (APS), and sample buffer were purchased from Bio-Rad (Hercules, CA, USA).
10
Oligonucleotide Synthesis and Purification
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The DNA oligonucleotides were synthesized and purified through HPLC by Sangon Biotech (Shanghai) Co., Ltd. (Shanghai, China). The sequences of the miRNA and DNA are given in Table S1 . The Endonuclease IV and 10×NEBuffer 3 (1000 mM NaCl, 500 mM Tris-HCl (pH 7.9), 100 mM MgCl2 and 10 mM DTT) were obtained from New England Biolabs (Ipswich, MA, USA). DSN and 10 × DSN master buffer (500 mM Tris-HCl, 50 mM MgCl2, 10 mM D, L-dithiothreitol (DTT), pH 8.0) were purchased from Evrogen (Moscow, Russia). The fetal bovine serum (FBS) was gained from Gibco (Carlsbad, CA, USA). The RNase inhibitor, DEPC-treated water, 10 × TBE buffer (225 mM Tris-Boric Acid, 50 mM EDTA, pH 8.0), 10 × phosphate-buffered saline (PBS), 4S green plus nucleic acid stain, and 10 × RNA glycerol gel loading buffer were obtained from Sangon Biological Engineering Technology & Services Co., Ltd. (Shanghai, China). Furthermore, 30% Acrylamide/bis-acrylamide, 29:1 (3.3% crosslinker), Ammonium Persulfate (APS), and N, N, N′, N′-Tetramethylethylenediamine (TEMED) were purchased from Bio-Rad Laboratories, Inc. (Hercules, CA, USA). Other chemicals were of analytical grade and were obtained from Sinopharm Chemical Reagents (Shanghai, China).
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