T

Tetrahydrobiopterin catalytic functions, 114-115 low-temperature optical absorption spectral detection, see Nitric oxide synthase

Tfam essentiality, 411

mitochondrial DNA maintenance, 411 tissue-specific knockout mouse generation cre-loxP recombination system, 412-414, 417, 420

embryonic stem cell culture and transfection, 418-419 flp-frt recombination system, 413, 420 mating of animals, 419-420 targeting vector construction homologous sequence in vector, 414 Lambda FIX II mouse genomic library screening, 415 phage DNA isolation, 415-416 Southern blot screening of homologous recombination, 414-415, 417 subcloning, 416-418 Thioredoxin enzyme-linked immunospot assay coating of plates, 27 development of plates, 30 materials, 25-27 mononuclear cell and monocytes isolation from human blood, 28-29 plating, 29 stimulation, 29 principles, 24-25

spot evaluation and enumeration, 30 statistical analysis, 30-31 functions, 23

oxidative stress response, 22 Thioredoxin reductase enzyme-linked immunospot assay coating of plates, 27 data presentation, 31-32 development of plates, 30 inducers of secretion, 31, 34-35 materials, 25-27 mononuclear cell and monocytes isolation from human blood, 28-29 plating, 29 stimulation, 29 principles, 24-25

spot evaluation and enumeration, 30 statistical analysis, 30-31 validation flow cytometry, 33-34 immunoprecipitation, 33 Western blot, 33 functions, 23

oxidative stress response, 22 radiolysis studies, 43-44 Topoisomerase I Iff expression during cell cycle, 448 functions, 448

HeLa cell cycle synchronization for studies,

449-450 messenger RNA

protein binding to 3'-untranslated region electrophoretic mobility shift assay,

454-456 redox sensitivity studies, 456-459 stability assay, 450 3'-untranslated region regulation of expression, reporter assay, 450, 452, 454

Transgenic mouse conditional knockout applications, 420-421

glutathione peroxidase, see Glutathione peroxidase heme oxygenase, see Heme oxygenase Huntington's disease studies, see Huntington's disease myocardial ischemia-reperfusion injury studies, see Myocardial ischemia-reperfusion injury superoxide dismutase, see Superoxide dismutase

Tfam tissue-specific knockout, see Tfam

Treponema pallidum culture overview, 140, 142 rabbit testis culture extraction media, 143-144 harvesting, 144-145 propagation, 144 SflEp cell coculture antioxidant effects, 147 historical perspective, 142-143 infection, 145 medium, 145

oxygen concentration effects, 146 quantitative analysis, 145-146 SflEp cell culture, 145 neelaredoxin, see Neelaredoxin oxygen sensitivity, 140-142, 155-156 redox protein homologs, 154-155

2,3,5-Triphenyl-2H-tetrazolium chloride, myocardial infarct staining, 334-336, 363-364

TTC, see 2,3,5-Triphenyl-2//-tetrazolium chloride

Two-hybrid assay, see Mammalian two-hybrid system; Yeast two-hybrid system

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