General procedure for preparing gel columns was followed. Briefly, 12% resolving gel was prepared by mixing 3.33 ml water, 2.5 mL Tris-HCl buffer (pH-8.8), 4 mL mixture of acrylamide and Bis-acrylamide (Bio-Rad) in ratio 29:1, 100 µL of 10% SDS, 60 µL Ammonium persulphate (APS) (Sigma Aldrich) and 6.5 µL of Tetramethylethylenediamine (TEMED) (Fisher Scientific). 6% stacking gel was prepared by adding 700 µL mixture of acrylamide: Bis-acrylamide in ratio 29:1, 675 µL of Tris-HCl buffer (pH-6.8) and 50 µL of 10% SDS. To this 25 µL of APS and 20 µL of TEMED were added. For capillary (Fisherbrand) preparation, resolving gel columns of 1.5 or 2.5 cm length were made by injecting 30 or 50 µL of resolving gel solution into the capillary tube, respectively. This was followed by injecting 10 µL of stacking gel solution to form stacking gel of 0.5 cm length. The standard PAGE slab was prepared using the above mentioned solutions to form resolving and stacking gels of 4 cm and 1 cm length.
Ammonium persulphate aps
Manufactured by Merck Group
Sourced in Germany, India
Ammonium persulphate (APS) is a chemical compound commonly used as a laboratory reagent. It is a crystalline, white, water-soluble solid with the chemical formula (NH4)2S2O8. APS is primarily used as an oxidizing agent and initiator in various chemical reactions and processes.
Automatically generated - may contain errors
Lab products found in correlation
Poloxamer 407, by Merck Group (2 mentions)
Nano ha, by Merck Group (2 mentions)
Bis acrylamide, by Bio-Rad (2 mentions)
Chitosan, by Merck Group (2 mentions)
Acrylic acid, by Merck Group (2 mentions)
Sodium chloride (nacl), by Carl Roth (2 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Tetramethylethylenediamine temed, by Thermo Fisher Scientific (1 mentions)
Caso4, by Merck Group (1 mentions)
Sodium alginate, by Merck Group (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by BD (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Nano3, by Merck Group (1 mentions)
Irgacure 2959, by Merck Group (1 mentions)
Spectra por, by Repligen (1 mentions)
N n methylenebis acrylamide mba, by Merck Group (1 mentions)
Theophylline, by Merck Group (1 mentions)
Carboxymethyl chitosan, by Merck Group (1 mentions)
N methylene bisacrylamide mba, by Merck Group (1 mentions)
15 protocols using ammonium persulphate aps
1
Capillary and Slab Gel Electrophoresis
2
Printable Calcium Phosphate Paste Formulation
3
Alginate/Chitosan Hydrogel Synthesis
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To create alginate/chitosan (SC) hydrogels, 2% sodium alginate (Aladdin, Shanghai, China) and 2% chitosan (Nantong Xingcheng Biological Industrial Co. Ltd., Nantong, China) (w/v) were first dissolved in MES (Sigma-Aldrich, St. Louis, MO, USA) buffer (pH 5.0) and stirred overnight until clean solutions were obtained. Precursor solution (10 mL) were mixed with 0.12 g 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), 0.12 g N-hydroxysuccinimide (NHS) and allowed to gel inside of a closed mold at room temperature overnight. To create alginate/chitosan/polyacrylamide (SCM) hydrogel (SCM4, SCM8, and SCM12), 2% sodium alginate and 2% chitosan were dissolved with 4%, 8%, or 12% of acrylamide (w/v), respectively. Additional 2% (w/v) N,N′-methylenebis(arylamide) (MBAA; Sigma-Aldrich) (12 μL), N,N,N′,N′-tetramethylethylenediamine (TEMED; Sigma-Aldrich) (2.7 μL), 0.27 M ammonium persulphate (APS; Sigma-Aldrich) (75 μL), and 0.75 M CaSO4 (Sigma-Aldrich) (64 μL) were syringe mixed with precursor solution containing sodium alginate/chitosan/4% acrylamide, along with EDC and NHS, to give SCM4 hydrogel for instance. sodium alginate (SA) hydrogels were prepared by mixing 2% sodium alginate solution (w/v) with 0.75 M CaSO4.
4
Hydrogel Synthesis and Characterization
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Acrylamide (AAm, BDH), sodium alginate (SA, Daejung Korea), 2-acrylamido-2-methylpropane sulfonic acid (AMPS, Alfa Aesar), N,N′-methylenebisacrylamide (MBA, BDH), ammonium persulphate (APS, Sigma Aldrich), phosphate buffers (pH = 3, 7 and 9, Daejung Korea), crystal violet (CV, Sigma Aldrich), NaNO3 (Merck), NaOH (BDH) and HCl (Sigma) reagents were used as received in this work. Milli-Q double deionized distilled water was used for all experimental work.
5
Antimicrobial Nanocomposite Hydrogel Synthesis
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The monomer (acrylamide, AAm), crosslinker (N,N-Methylenebis(acrylamide), MBA), and carbon nanospecies (carboxyl-functionalized multiwall carbon nanotubes, CNTs) were purchased from Sigma. Polyacrylamide (PAAm), with an average molecular weight of 3.76 × 105 g/mol, was obtained in the laboratory through the free radical photopolymerization of the monomer in an aqueous solution in the presence of Irgacure 2959 (Sigma, St. Louis, MO, USA), purified through dialysis against distilled water (dH2O) using dialysis membranes (SpectraPor, MWCO 12–14 kDa, Repligen, CA, USA). The polymer’s synthesis and the method employed for the assessment of its viscosity and average molecular weight are described in Appendix A . Ammonium persulphate (APS, Sigma) and trietanolamine (TEA, Sigma) were used as the redox initiating system of the AAm-MBA system. Phosphate-buffered saline (PBS 0.01 M, pH 7.4, Sigma-Aldrich, St. Louis, MO, USA) was prepared according to manufacturer’s instructions. For the antimicrobial tests, Staphylococcus aureus (ATCC 6538TM) and Escherichia coli (ATCC 10536TM) prepared according to the manufacturer’s recommendations were used as references. Casein soyabean digest broth (TSB, Sigma) was used as dispersant for the lyophilized bacteria, while casein soyabean digest agar (TSA, Sigma) was used as culture medium.
6
Isolation of Cellulose Nanocrystals (CNC)
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Sodium chlorite (NaClO2), sodium hydroxide (NaOH), sulphuric acid (H2SO4) and glacial acetic acid (99%) were used for the isolation of CNC. Acrylic acid (99%) was purchased from Sigma Aldrich (Darmstadt, Germany). Laboratory-grade N,N′-methylenebis(acrylamide) (MBA, 99%), ammonium persulphate (APS, 99%), and theophylline (>99%) were purchased from Sigma-Aldrich (Darmstadt, Germany) for use as the cross-linker, initiator, and drug, respectively.
7
Printable Calcium Phosphate Paste Formulation
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PCaP paste was prepared by mixing a powder particle phase and a liquid
phase. Briefly, the optimal distribution of the particles and liquid phases that
allowed the paste to be printable was 70% and 30% w/w, respectively. The powder
consisted of microparticles of milled α-TCP (average size 3.37
µm, Cambioceramics, The Netherlands) mixed with 4% w/w nano-HA (average size 200
nm, Sigma-Aldrich). The liquid phase consisted of a shear-thinning hydrogel
precursor solution dissolved in phosphate-buffered saline (PBS), supplemented
with ammonium persulphate (APS, Sigma Aldrich, 25 mM), to form a 40% (w/v)
solution. The dissolved polymer, forming this hydrogel precursor, consisted of a
biodegradable and crosslinkable poloxamer derivative (P-CL-MA), which was custom
synthesized by grafting a biodegradable ∊-caprolactone ester
block and a methacrylate group onto both terminal hydroxyl groups of poloxamer
407 (Sigma-Aldrich), as previously described.[24 ] Before mixing, the powder and liquid phases
were stored separately at 4 °C for 30 min and finally the P-CL-MA solution was
added to a composite solid particle at 4 °C and manually mixed with a spatula.
To ensure homogeneous distribution of solid particles, the mixing process was
performed for 3 min at 4 °C. Finally, the PCaP paste was loaded into a 5 mL
dispensing cartridge, closed with retainer caps, and stored at 4 °C until
using.
phase. Briefly, the optimal distribution of the particles and liquid phases that
allowed the paste to be printable was 70% and 30% w/w, respectively. The powder
consisted of microparticles of milled α-TCP (average size 3.37
µm, Cambioceramics, The Netherlands) mixed with 4% w/w nano-HA (average size 200
nm, Sigma-Aldrich). The liquid phase consisted of a shear-thinning hydrogel
precursor solution dissolved in phosphate-buffered saline (PBS), supplemented
with ammonium persulphate (APS, Sigma Aldrich, 25 mM), to form a 40% (w/v)
solution. The dissolved polymer, forming this hydrogel precursor, consisted of a
biodegradable and crosslinkable poloxamer derivative (P-CL-MA), which was custom
synthesized by grafting a biodegradable ∊-caprolactone ester
block and a methacrylate group onto both terminal hydroxyl groups of poloxamer
407 (Sigma-Aldrich), as previously described.[24 ] Before mixing, the powder and liquid phases
were stored separately at 4 °C for 30 min and finally the P-CL-MA solution was
added to a composite solid particle at 4 °C and manually mixed with a spatula.
To ensure homogeneous distribution of solid particles, the mixing process was
performed for 3 min at 4 °C. Finally, the PCaP paste was loaded into a 5 mL
dispensing cartridge, closed with retainer caps, and stored at 4 °C until
using.
8
Docetaxel Loaded Hydrogel Formulation
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Docetaxel was purchased from Xian Wharton Bio-Tech Co., Ltd., Caotan Rd Xi’an, China. Carboxymethyl chitosan (CMC), β-cyclodextrin(βCD), 2-Acrylamido-2-methylpropane sulfonic acid (AMPS), N, N′-Methylene bis(acrylamide) (MBA) and Ammonium per sulphate (APS) were supplied by Sigma-Aldrich, St. Louis, MO, United States of America. All aqueous solutions were prepared with deionized water obtained from Pharmaceutics Research Lab of the Islamia University of Bahawalpur, Bahawalpur, Pakistan.
9
Protein Purification and SDS-PAGE Analysis
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The purified protein sample was estimated to be 90% pure, based on SDS-PAGE gels. Tris buffer, sodium cacodylate buffer, LB broth, kanamycin, isopropyl-ß-D-thiogalacto-pyranoside (IPTG), imidazole, sodium dodecylsulphate (SDS), NaCl, acrylamide/bisacrylamide solution 29:1, (30% w/v), N,N,N′,N′-tetramethylethylenediamine (TEMED), ammonium peroxide, Rotigarose His/Ni beads, methanol, glycerol, glycine, sodium chloride, powdered milk, and acetic acid were acquired from Carl Roth (Karlsruhe, Germany). Amonium-acetate buffer, ammonium persulphate (APS), β-mercaptoethanol, Tween 20, bromphenol blue, MOPS buffer, and diphenylamine (DPA) were acquired from Sigma-Aldrich (St. Louis, MO, USA). The Arg2-2-naphthylamide (Arg-Arg–2NA) was produced by Bachem (Bubendorf, Switzerland); 2-naphthylamine (2-NA), 2-mercaptoethanol, and DNase I from bovine pancreas were produced by Merck (Darmstadt, Germany). For visualization of SDS-PAGE gels, we used PhastGel Blue R tablets (Pharmacia, Uppsala, Sweden). The EDTA was obtained from Kemika (Zagreb, Croatia), and metal (zinc, copper, cobalt, manganese) standard nitrate solutions were produced by Merck (Darmstadt, Germany).
10
Synthesis of Polymer Latexes via Emulsion Polymerization
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Technical grade monomers methyl methacrylate (MMA, 98% purity, Quimidroga, Barcelona, Spain), butyl acrylate (BA, 98% purity, Quimidroga), methacrylic acid (MAA, 99% purity, Sigma-Aldrich, Taufkirchen, Germany) and itaconic acid (ITA, 99% purity, Sigma-Aldrich) were used to synthesize the latexes. Polyethyleneglycol methacrylate (PEGMA, 5000 g/mol, 99% purity, Evonik, Essen, Germany) was used as macromonomer. Sodium dodecyl sulphate (SDS, 99% purity, Sigma-Aldrich) was used as surfactant to stabilize the droplets and ammonium persulphate (APS, 99% purity, Sigma-Aldrich) to initiate the polymerization. All the chemicals were used as received. Deionized MilliQ water was used as polymerization media.
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