Variant Assay:Article Title: Nucleotide and metalloid-driven conformational changes in the arsenite efflux ATPase ArsA
Article Snippet: .. The reaction mixture (100 μL) consisted of 50 mM HEPES pH 7.5, containing 5 mM MgCl 2 , 2 mM phosphoenol pyruvate (Roche), 20 U/mL pyruvate kinase from rabbit muscle (MP Biomedicals), 20 U/mL L-lactate dehydrogenase from rabbit muscle (Sigma), 0.2 mM NADH (Roche), 2-5 μM ArsA or its variant and varying concentrations of sodium arsenite (0, 0.05, 0.1, 0.2, 0.5 and 0.8 mM). ..
Incubation:Article Title: Ubiquitin-Specific Protease 2 in the Ventromedial Hypothalamus Modifies Blood Glucose Levels by Controlling Sympathetic Nervous Activation
Article Snippet: .. Mitochondria were incubated with the following (in mM): 50 4-(2-hydroxyethyl) 1-piperazineethanesulfonic acid, 3 magnesium chloride, 50 potassium cyanide, 0.2 EDTA, 2 phosphoenolpyruvic acid, and 100 mM rotenone, 10 units/ml lactate dehydrogenase (Oriental Yeast), and 10 units/ml pyruvate kinase (MP Biomedicals) for 5min, and then treated with 2 mM ATP and 500 mM NADH in the presence or absence of 10mg/ml oligomycin (Fujifilm Wako Pure Chemical). ..
Inhibition:Article Title: Phylogenetic Investigation of the 100 kDa Hexokinase Enzyme Family with the Topiary Ancestral Sequence Reconstruction Pipeline
Article Snippet: .. Inhibition assays were conducted by coupling the production of ADP to the oxidation of NADH via an enzyme-coupled spectrophotometric assay using rabbit muscle pyruvate kinase (MP Biomedicals) and rabbit muscle or porcine heart lactate dehydrogenase (MilliporeSigma). .. Reaction progress was followed at 340 nm at 25 °C (Agilent Cary UV-Vis spectrophotometer).
Spectrophotometric Assay:Article Title: Phylogenetic Investigation of the 100 kDa Hexokinase Enzyme Family with the Topiary Ancestral Sequence Reconstruction Pipeline
Article Snippet: .. Inhibition assays were conducted by coupling the production of ADP to the oxidation of NADH via an enzyme-coupled spectrophotometric assay using rabbit muscle pyruvate kinase (MP Biomedicals) and rabbit muscle or porcine heart lactate dehydrogenase (MilliporeSigma). .. Reaction progress was followed at 340 nm at 25 °C (Agilent Cary UV-Vis spectrophotometer).
Staining:Article Title: Altered Mitochondrial Morphology and Bioenergetics in a New Yeast Model Expressing Aβ42.
Article Snippet: .. 2023, 24, 900 13 of 20 ADP, ampicillin, Anti-Rabbit IgG Peroxidase antibody, antimycin A, ATP, 3-amino-1,2,4triazole, carbonyl cyanide m-chlorophenylhydrazon (CCCP), DABCO, P5-di(adenosine5)pentaphosphate (Ap5A), EDTA, EGTA, fatty acid-free BSA, glucose, glucose-6-phosphate dehydrogenase, LiAc, mannitol, MgCl2, Mowiol 4-88, NaCl, NADP, (NH4)2SO4, oligomycin, Phenol Red, phosphoenolpyruvate, pyruvate kinase, rotenone, succinic acid, and tert-butyl hydroperoxide were from Sigma-Aldrich (St. Louis, MO, USA); Coomassie G-250 and zymolyase were from MP Biomedicals (Santa Ana, CA, USA); CaCl2, K2HPO4, KCl, KH2PO4, NaCl, and safranin O were from Merck (Darmstadt, Germany); 10× DNA Loading Dye, 10× G+ buffer, 10× O+ buffer, 10× R+ buffer, BSA, dihydroetidium, Sytox Green Dead Cell Stain, DMSO, Gene Jet Gel Extraction Kit, Gene Jet Plasmid Miniprep Kit, Gene Ruler 100 bp+, Gene Ruler 1 kb, Glycogen, Mitotracker Red CmxRos, NotI restriction endonuclease, PageRulerTM Prestained Protein Ladder, Phusion High-Fidelity PCR Kit, PvuII restriction endonuclease, Rapid DNA Ligation Kit, RNAse-A, SuperSignalTM West Dura Extended Duration Substrate, and XhoI restriction endonuclease were from Thermo Fisher Scientific (Waltham, MA, USA); Agar, agarose LE2, ethidium bromide, and glycerol (ultra-pure) were from Helicon (Moscow, Russian Federation); LB BROTH Miller (Luria–Bertani) and NaOAc were from Amresco (Dallas, TX, USA); BbsI (BpiI) restriction endonuclease was from New England Biolabs (Ipswich, MA, USA); sorbitol was from Dia-M (Moscow, Russia); oligonucleotides were from DNA-Synthesis (Moscow, Russia). ..
Gel Extraction:Article Title: Altered Mitochondrial Morphology and Bioenergetics in a New Yeast Model Expressing Aβ42.
Article Snippet: .. 2023, 24, 900 13 of 20 ADP, ampicillin, Anti-Rabbit IgG Peroxidase antibody, antimycin A, ATP, 3-amino-1,2,4triazole, carbonyl cyanide m-chlorophenylhydrazon (CCCP), DABCO, P5-di(adenosine5)pentaphosphate (Ap5A), EDTA, EGTA, fatty acid-free BSA, glucose, glucose-6-phosphate dehydrogenase, LiAc, mannitol, MgCl2, Mowiol 4-88, NaCl, NADP, (NH4)2SO4, oligomycin, Phenol Red, phosphoenolpyruvate, pyruvate kinase, rotenone, succinic acid, and tert-butyl hydroperoxide were from Sigma-Aldrich (St. Louis, MO, USA); Coomassie G-250 and zymolyase were from MP Biomedicals (Santa Ana, CA, USA); CaCl2, K2HPO4, KCl, KH2PO4, NaCl, and safranin O were from Merck (Darmstadt, Germany); 10× DNA Loading Dye, 10× G+ buffer, 10× O+ buffer, 10× R+ buffer, BSA, dihydroetidium, Sytox Green Dead Cell Stain, DMSO, Gene Jet Gel Extraction Kit, Gene Jet Plasmid Miniprep Kit, Gene Ruler 100 bp+, Gene Ruler 1 kb, Glycogen, Mitotracker Red CmxRos, NotI restriction endonuclease, PageRulerTM Prestained Protein Ladder, Phusion High-Fidelity PCR Kit, PvuII restriction endonuclease, Rapid DNA Ligation Kit, RNAse-A, SuperSignalTM West Dura Extended Duration Substrate, and XhoI restriction endonuclease were from Thermo Fisher Scientific (Waltham, MA, USA); Agar, agarose LE2, ethidium bromide, and glycerol (ultra-pure) were from Helicon (Moscow, Russian Federation); LB BROTH Miller (Luria–Bertani) and NaOAc were from Amresco (Dallas, TX, USA); BbsI (BpiI) restriction endonuclease was from New England Biolabs (Ipswich, MA, USA); sorbitol was from Dia-M (Moscow, Russia); oligonucleotides were from DNA-Synthesis (Moscow, Russia). ..
Plasmid Preparation:Article Title: Altered Mitochondrial Morphology and Bioenergetics in a New Yeast Model Expressing Aβ42.
Article Snippet: .. 2023, 24, 900 13 of 20 ADP, ampicillin, Anti-Rabbit IgG Peroxidase antibody, antimycin A, ATP, 3-amino-1,2,4triazole, carbonyl cyanide m-chlorophenylhydrazon (CCCP), DABCO, P5-di(adenosine5)pentaphosphate (Ap5A), EDTA, EGTA, fatty acid-free BSA, glucose, glucose-6-phosphate dehydrogenase, LiAc, mannitol, MgCl2, Mowiol 4-88, NaCl, NADP, (NH4)2SO4, oligomycin, Phenol Red, phosphoenolpyruvate, pyruvate kinase, rotenone, succinic acid, and tert-butyl hydroperoxide were from Sigma-Aldrich (St. Louis, MO, USA); Coomassie G-250 and zymolyase were from MP Biomedicals (Santa Ana, CA, USA); CaCl2, K2HPO4, KCl, KH2PO4, NaCl, and safranin O were from Merck (Darmstadt, Germany); 10× DNA Loading Dye, 10× G+ buffer, 10× O+ buffer, 10× R+ buffer, BSA, dihydroetidium, Sytox Green Dead Cell Stain, DMSO, Gene Jet Gel Extraction Kit, Gene Jet Plasmid Miniprep Kit, Gene Ruler 100 bp+, Gene Ruler 1 kb, Glycogen, Mitotracker Red CmxRos, NotI restriction endonuclease, PageRulerTM Prestained Protein Ladder, Phusion High-Fidelity PCR Kit, PvuII restriction endonuclease, Rapid DNA Ligation Kit, RNAse-A, SuperSignalTM West Dura Extended Duration Substrate, and XhoI restriction endonuclease were from Thermo Fisher Scientific (Waltham, MA, USA); Agar, agarose LE2, ethidium bromide, and glycerol (ultra-pure) were from Helicon (Moscow, Russian Federation); LB BROTH Miller (Luria–Bertani) and NaOAc were from Amresco (Dallas, TX, USA); BbsI (BpiI) restriction endonuclease was from New England Biolabs (Ipswich, MA, USA); sorbitol was from Dia-M (Moscow, Russia); oligonucleotides were from DNA-Synthesis (Moscow, Russia). ..
Polymerase Chain Reaction:Article Title: Altered Mitochondrial Morphology and Bioenergetics in a New Yeast Model Expressing Aβ42.
Article Snippet: .. 2023, 24, 900 13 of 20 ADP, ampicillin, Anti-Rabbit IgG Peroxidase antibody, antimycin A, ATP, 3-amino-1,2,4triazole, carbonyl cyanide m-chlorophenylhydrazon (CCCP), DABCO, P5-di(adenosine5)pentaphosphate (Ap5A), EDTA, EGTA, fatty acid-free BSA, glucose, glucose-6-phosphate dehydrogenase, LiAc, mannitol, MgCl2, Mowiol 4-88, NaCl, NADP, (NH4)2SO4, oligomycin, Phenol Red, phosphoenolpyruvate, pyruvate kinase, rotenone, succinic acid, and tert-butyl hydroperoxide were from Sigma-Aldrich (St. Louis, MO, USA); Coomassie G-250 and zymolyase were from MP Biomedicals (Santa Ana, CA, USA); CaCl2, K2HPO4, KCl, KH2PO4, NaCl, and safranin O were from Merck (Darmstadt, Germany); 10× DNA Loading Dye, 10× G+ buffer, 10× O+ buffer, 10× R+ buffer, BSA, dihydroetidium, Sytox Green Dead Cell Stain, DMSO, Gene Jet Gel Extraction Kit, Gene Jet Plasmid Miniprep Kit, Gene Ruler 100 bp+, Gene Ruler 1 kb, Glycogen, Mitotracker Red CmxRos, NotI restriction endonuclease, PageRulerTM Prestained Protein Ladder, Phusion High-Fidelity PCR Kit, PvuII restriction endonuclease, Rapid DNA Ligation Kit, RNAse-A, SuperSignalTM West Dura Extended Duration Substrate, and XhoI restriction endonuclease were from Thermo Fisher Scientific (Waltham, MA, USA); Agar, agarose LE2, ethidium bromide, and glycerol (ultra-pure) were from Helicon (Moscow, Russian Federation); LB BROTH Miller (Luria–Bertani) and NaOAc were from Amresco (Dallas, TX, USA); BbsI (BpiI) restriction endonuclease was from New England Biolabs (Ipswich, MA, USA); sorbitol was from Dia-M (Moscow, Russia); oligonucleotides were from DNA-Synthesis (Moscow, Russia). ..
DNA Ligation:Article Title: Altered Mitochondrial Morphology and Bioenergetics in a New Yeast Model Expressing Aβ42.
Article Snippet: .. 2023, 24, 900 13 of 20 ADP, ampicillin, Anti-Rabbit IgG Peroxidase antibody, antimycin A, ATP, 3-amino-1,2,4triazole, carbonyl cyanide m-chlorophenylhydrazon (CCCP), DABCO, P5-di(adenosine5)pentaphosphate (Ap5A), EDTA, EGTA, fatty acid-free BSA, glucose, glucose-6-phosphate dehydrogenase, LiAc, mannitol, MgCl2, Mowiol 4-88, NaCl, NADP, (NH4)2SO4, oligomycin, Phenol Red, phosphoenolpyruvate, pyruvate kinase, rotenone, succinic acid, and tert-butyl hydroperoxide were from Sigma-Aldrich (St. Louis, MO, USA); Coomassie G-250 and zymolyase were from MP Biomedicals (Santa Ana, CA, USA); CaCl2, K2HPO4, KCl, KH2PO4, NaCl, and safranin O were from Merck (Darmstadt, Germany); 10× DNA Loading Dye, 10× G+ buffer, 10× O+ buffer, 10× R+ buffer, BSA, dihydroetidium, Sytox Green Dead Cell Stain, DMSO, Gene Jet Gel Extraction Kit, Gene Jet Plasmid Miniprep Kit, Gene Ruler 100 bp+, Gene Ruler 1 kb, Glycogen, Mitotracker Red CmxRos, NotI restriction endonuclease, PageRulerTM Prestained Protein Ladder, Phusion High-Fidelity PCR Kit, PvuII restriction endonuclease, Rapid DNA Ligation Kit, RNAse-A, SuperSignalTM West Dura Extended Duration Substrate, and XhoI restriction endonuclease were from Thermo Fisher Scientific (Waltham, MA, USA); Agar, agarose LE2, ethidium bromide, and glycerol (ultra-pure) were from Helicon (Moscow, Russian Federation); LB BROTH Miller (Luria–Bertani) and NaOAc were from Amresco (Dallas, TX, USA); BbsI (BpiI) restriction endonuclease was from New England Biolabs (Ipswich, MA, USA); sorbitol was from Dia-M (Moscow, Russia); oligonucleotides were from DNA-Synthesis (Moscow, Russia). ..
DNA Synthesis:Article Title: Altered Mitochondrial Morphology and Bioenergetics in a New Yeast Model Expressing Aβ42.
Article Snippet: .. 2023, 24, 900 13 of 20 ADP, ampicillin, Anti-Rabbit IgG Peroxidase antibody, antimycin A, ATP, 3-amino-1,2,4triazole, carbonyl cyanide m-chlorophenylhydrazon (CCCP), DABCO, P5-di(adenosine5)pentaphosphate (Ap5A), EDTA, EGTA, fatty acid-free BSA, glucose, glucose-6-phosphate dehydrogenase, LiAc, mannitol, MgCl2, Mowiol 4-88, NaCl, NADP, (NH4)2SO4, oligomycin, Phenol Red, phosphoenolpyruvate, pyruvate kinase, rotenone, succinic acid, and tert-butyl hydroperoxide were from Sigma-Aldrich (St. Louis, MO, USA); Coomassie G-250 and zymolyase were from MP Biomedicals (Santa Ana, CA, USA); CaCl2, K2HPO4, KCl, KH2PO4, NaCl, and safranin O were from Merck (Darmstadt, Germany); 10× DNA Loading Dye, 10× G+ buffer, 10× O+ buffer, 10× R+ buffer, BSA, dihydroetidium, Sytox Green Dead Cell Stain, DMSO, Gene Jet Gel Extraction Kit, Gene Jet Plasmid Miniprep Kit, Gene Ruler 100 bp+, Gene Ruler 1 kb, Glycogen, Mitotracker Red CmxRos, NotI restriction endonuclease, PageRulerTM Prestained Protein Ladder, Phusion High-Fidelity PCR Kit, PvuII restriction endonuclease, Rapid DNA Ligation Kit, RNAse-A, SuperSignalTM West Dura Extended Duration Substrate, and XhoI restriction endonuclease were from Thermo Fisher Scientific (Waltham, MA, USA); Agar, agarose LE2, ethidium bromide, and glycerol (ultra-pure) were from Helicon (Moscow, Russian Federation); LB BROTH Miller (Luria–Bertani) and NaOAc were from Amresco (Dallas, TX, USA); BbsI (BpiI) restriction endonuclease was from New England Biolabs (Ipswich, MA, USA); sorbitol was from Dia-M (Moscow, Russia); oligonucleotides were from DNA-Synthesis (Moscow, Russia). ..
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