Hydrogen tetrachloroauratetrihydrate (HAuCl4·3H2O) was purchased from Millipore Sigma (Shanghai, China). Nitrocellulose membranes (Hi-Flow plus 120, Merck Millipore), sample pad (cellulose fiber, Merck Millipore), conjugate pad (cellulose fiber, Merck Millipore), absorbent pads, adhesive backing cards and goat anti-mouse IgG were obtained from Rojan Azma Co (Tehran, Iran). Trisodium citrate dehydrate, bovine serum albumin, Tween-20, phosphate buffer solution, PEG20000, glucose and 0.22 μm filters were purchased from Merck Millipore (Burlington, MA, USA). Gliadin Monoclonal Antibody (14D5) was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
Peg20000
Manufactured by Merck Group
Sourced in United States
PEG20000 is a polyethylene glycol (PEG) compound with an average molecular weight of 20,000 Daltons. It is a water-soluble and non-toxic polymer commonly used in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (3 mentions)
Peg 8000, by Merck Group (2 mentions)
Peg 200, by Merck Group (2 mentions)
Phosphate buffer solution (pbs), by Merck Group (1 mentions)
Trisodium citrate dehydrate, by Merck Group (1 mentions)
Glucose, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
0.22 μm filter, by Merck Group (1 mentions)
Yeast enolase, by Merck Group (1 mentions)
Monosodium phosphoenolpyruvate, by Merck Group (1 mentions)
Fructose 6 phosphate disodium salt, by Thermo Fisher Scientific (1 mentions)
Sodium chloride (nacl), by AppliChem (1 mentions)
Synergy 2 multi mode reader, by Agilent Technologies (1 mentions)
Adp glo kinase assay, by Promega (1 mentions)
Temozolomide, by Merck Group (1 mentions)
Peg 10 000, by Merck Group (1 mentions)
Ethylene glycol, by Merck Group (1 mentions)
Peg400, by Merck Group (1 mentions)
Nitrocellulose membrane she 1350225, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
17 protocols using peg20000
1
Rapid Gliadin Immunoassay Development
2
Enzymatic Assay for Yeast Enolase
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Yeast enolase, monosodium phosphoenolpyruvate, phosphoglucose isomerase type III from baker’s yeast and bovine serum albumin (BSA) were purchased from Sigma Aldrich (Sigma-Aldrich Chemie GmbH, Steinheim, Germany), fructose 6-phosphate disodium salt from Alfa Aesar (Thermo Fisher (Kandel) GmbH, Kandel, Germany), MOPS from AppliChem (AppliChem GmbH, Darmstadt, Germany), sodium chloride from CHEMSOLUTE (Th. Geyer GmbH & Co. KG, Renningen, Germany), polyethylene glycol with an average molar mass of 20,000 g mol−1 (PEG 20,000) from Merck (Merck KGaA, Darmstadt, Germany), polyethylene glycol 6000 from Serva (SERVA Electrophoresis GmbH, Heidelberg, Germany), magnesium chloride hexahydrate and sodium hydroxide from Roth (Bernd Kraft, Duisburg, Germany). An overview of all used chemicals, the CAS-numbers and purity is given in Table S1 in the supplementary materials .
3
Kinase Inhibition Assay for c-Met
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Active, recombinant c-Met enzyme (3 µg/mL, ProQinase, cat. no.: 0171-0000-1, Lot: 012) was incubated with ATP (3.1 µM (KM[ATP]), Promega, cat. no.: V6930), peptide substrate (0,25 mg/mL, poly GT, Merck Millipore, cat. no.: PO275), and compounds at the indicated concentrations. Reaction buffer was prepared according to ProQinase’s recommendation [60 mM HEPES, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na3VO4, 1.2 mM DTT (dithiothreitol), and 50 µg/mL PEG20,000, (reagents were obtained from Merck Millipore)]. Enzyme activity was assayed in 384 well, low-volume white microtiter plates (Corning®, Merck Millipore, cat. no.: CLS4513-50EA). Compounds were preincubated with c-Met kinase for 30 min at room temperature and reactions were started by adding the substrate–ATP mixture. Reaction time was 60 min at room temperature. Enzyme reaction was stopped using ADP-Glo™ Kinase Assay (Promega, cat. no.: V6930) according to the manufacturer’s instructions (40 min incubation with ADP-Glo™ Reagent, 30 min incubation with Kinase detection reagent at 25 °C). All measurements were performed using a Synergy 2 Multi-Mode Reader (BioTek, Winooski, VT, USA). Raw enzyme activity values were converted to relative activity data using positive and negative controls. Determination of IC50 values was performed using GraphPad Prism 5.02 software.
4
Synthesis of PEGylated Paclitaxel and Temozolomide
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All chemicals including different types of PEG (ethylene glycol -Cat# 102466, PEG 200 -Cat# P3015, PEG 400 -Cat# P3265, PEG 8000 -Cat# 89510, PEG 10 000 -Cat# 81280 and PEG 20 000 -Cat# 95172 -all bis-hydroxy terminated) and glucono-d-lactone were purchased from Sigma Aldrich, and used as received. MilliQ Water (resistivity 18.2 MO) was used throughout the study. Paclitaxel (Taxol s ) was obtained from LC Laboratories USA and Temozolomide from Sigma Aldrich. The solution phase synthesis of 1 was adapted from the methods (ESI †) previously reported by Ko ¨nig and Ro ¨dela 48 and Gagnon et al.
5
Lanthanide-Doped Upconversion Phosphor Protocol
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UCP (NaYF4:Yb3+,Er3+) with excitation and emission
spectrum peaks of 980 and 541.5 nm, respectively, was prepared by
Dr. Yan Zheng from Shanghai Kerune Phosphor Technology Co., Ltd. (Shanghai,
China). Nitrocellulose membrane (SHE 1350225) and glass fiber (GFCP20300) were
purchased from Millipore Corp. (Bedford, MA, USA). Absorbent papers (Nos 470 and
903) were obtained from Schleicher & Schuell, Inc. (Keene, NH, USA).
Plastic cartridges were designed by our group and processed by Shenzhen
Jincanhua Industry Co. (Shenzhen, China).
Formaldehyde, HCl, NaOH, KCl, NaCl, PEG20000, glycerin, bovine serum albumin V
(BSA), and casein were of analytical grade and purchased from
Sigma–Aldrich (St. Louis, MO, USA). The real samples, including
flour, fruit juice, gourmet powder, milk powder, putty powder, and sucrose, were
all obtained from the local market, and soil samples were excavated from a
parterre. Viscera obtained from Balb/C mice, including heart, liver, lung, and
spleen, were divided into two parts. One part was stored at
−20 °C as fresh specimen, and the other part
was incubated at 37 °C for two weeks as decomposed
specimen.
spectrum peaks of 980 and 541.5 nm, respectively, was prepared by
Dr. Yan Zheng from Shanghai Kerune Phosphor Technology Co., Ltd. (Shanghai,
China). Nitrocellulose membrane (SHE 1350225) and glass fiber (GFCP20300) were
purchased from Millipore Corp. (Bedford, MA, USA). Absorbent papers (Nos 470 and
903) were obtained from Schleicher & Schuell, Inc. (Keene, NH, USA).
Plastic cartridges were designed by our group and processed by Shenzhen
Jincanhua Industry Co. (Shenzhen, China).
Formaldehyde, HCl, NaOH, KCl, NaCl, PEG20000, glycerin, bovine serum albumin V
(BSA), and casein were of analytical grade and purchased from
Sigma–Aldrich (St. Louis, MO, USA). The real samples, including
flour, fruit juice, gourmet powder, milk powder, putty powder, and sucrose, were
all obtained from the local market, and soil samples were excavated from a
parterre. Viscera obtained from Balb/C mice, including heart, liver, lung, and
spleen, were divided into two parts. One part was stored at
−20 °C as fresh specimen, and the other part
was incubated at 37 °C for two weeks as decomposed
specimen.
6
Enzymatic DNA Modification Reagents
7
Functionalization of Micro- and Nanoparticles
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PEG 20000, PBS solution, and 11-mercaptohexanoic acid were purchased from Sigma-Aldrich. PS microparticles (2.8, 2.2, 1, and 0.5 μm) with streptavidin ligands, 40-nm yellow-green fluorescent (505/515) nanobeads with biotin ligands, and 2-μm silica particles were purchased from Thermo Fisher Scientific. PS particles (500 and 300 nm) were purchased from Bangs Laboratories.
8
Edible Sunflower Oil Formulations
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Edible sunflower oil (Zvijezda plus d.o.o., Zagreb, Croatia) was purchased from a local supermarket. Polyethylene glycols with average molecular weights of 1500 g/mol (PEG 1500) and 6000 g/mol (PEG 6000) were purchased from Acros Organics (Geel, Belgium), while the 20,000 g/mol polyethylene glycol (PEG 20,000) was obtained from Sigma-Aldrich (Taufkirchen, Germany). Dried mint leaves (Mentha piperita L.) were purchased from Suban, Croatia. Plant materials were collected during the flowering season of 2019 in the north-western part of Croatia, dried naturally, and stored in ambient conditions before use.
9
Synthesis and Characterization of Cu-Doped Titania Nanocomposites
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Titanium isopropoxide (TIP), ZrO(NO3)2.xH2O, Ca(NO3)2.4H2O, Cu(NO3)2.3H2O, acetic acid, nitric acid, and all commercially available solvents were purchased from Merck Company and used without further purification. Polyethylene glycol (PEG 20000) was purchased from Sigma-Aldrich.
X-ray diffraction (XRD) pattern was acquired by STOE X-Ray Diffractometer Tube Anode: Cu Wavelength 1.5406 A° (Cu Kα, Filter Ni). Scanning electron microscopy (SEM) was carried out on Philips XL30. A Varian (AA220) flame atomic absorption spectrometer (AAS) (air/acetylene flame) was used for copper ion determinations. The specific surface area was determined by the Brunauer-Emmett-Teller method (BET) using Micromeritics Instrument Corporation TriStar II 3020 V1.03 (V1.03), and the pore volume and pore size distribution were calculated from the desorption branch of the isotherm as measured by the Barrett-Joyner-Halenda method (BJH). Thermogravimetric analyses (TGA) were recorded from room temperature to 800 °C under a nitrogen atmosphere by a TGA (TA Q600) thermogravimetric analyzer. Temperature programmed reduction (TPR) and temperature-programmed desorption (TPD) experiments were carried out using a BELCAT-An instrument equipped with a thermal conductivity detector.
X-ray diffraction (XRD) pattern was acquired by STOE X-Ray Diffractometer Tube Anode: Cu Wavelength 1.5406 A° (Cu Kα, Filter Ni). Scanning electron microscopy (SEM) was carried out on Philips XL30. A Varian (AA220) flame atomic absorption spectrometer (AAS) (air/acetylene flame) was used for copper ion determinations. The specific surface area was determined by the Brunauer-Emmett-Teller method (BET) using Micromeritics Instrument Corporation TriStar II 3020 V1.03 (V1.03), and the pore volume and pore size distribution were calculated from the desorption branch of the isotherm as measured by the Barrett-Joyner-Halenda method (BJH). Thermogravimetric analyses (TGA) were recorded from room temperature to 800 °C under a nitrogen atmosphere by a TGA (TA Q600) thermogravimetric analyzer. Temperature programmed reduction (TPR) and temperature-programmed desorption (TPD) experiments were carried out using a BELCAT-An instrument equipped with a thermal conductivity detector.
10
In Vitro Kinase Assay of Neuroligin-1
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To confirm ourselves the ability of some of those kinases to phosphorylate Nlg1, we used GST-Nlg1 and commercial tyrosine kinases. The in vitro kinase assay was performed by incubating a mix of purified GST-Nlg1 protein with the purified enzyme at 30 °C for 1 h, on a shaker. The mixture consisted of: (1) the purified protein: GST-Nlg1-WT (120 µg/ml) or GST-Nlg1-Y782A (120 µg/ml) or TrkC positive substrate, RBER-NTRK3tide (ProQinase, 25 µg/ml); (2) the enzyme at 1 µg/ml: GST-FGFR1 (V561M) (Sigma-Aldrich,), GST-TrkB or -TrkC (ProQinase GmbH); (3) buffer containing: 60 mM HEPES-NaOH, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, 0,005% PEG 20000, 50 µM ATP (Sigma, A2383). After incubation, 10 µl of 6× loading buffer (350 mM Tris-HCl, 10% SDS, 30% glycerol, 5% β-mercaptoethanol, 0.06% bromophenol blue, pH = 6.8) was added to each sample, and the mix was heated for 5 min at 60 °C. Samples (13 µl) of the resulting solution were loaded in polyacrylamide gels.
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