By Donald Armstrong
In Oxidants and Antioxidants: Ultrastructural and Molecular Biology Protocols, a panel of global professionals current state of the art molecular and ultrastructure strategies for investigating the pathophysiology of oxidative tension. defined in step by step aspect, those comfortably reproducible tools use the newest in immunodetection, X-ray diffraction, confocal microscopy, in situ hybridization, chemiluminescence, and cytofluorescence. extra suggestions reap the benefits of electron paramagnetic resonance, complex glycation endproducts, Western blotting research, transcription elements, p47 phox translocation, and microsphere supply platforms. spouse volumes, Oxidative rigidity Biomarkers and Antioxidant Protocols (2002), and unfastened Radical and Antioxidant Protocols (1998), specialise in the similar biochemical and mobile organic tools.
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Extra resources for Oxidants and Antioxidants: Ultrastructure and Molecular Biology Protocols
42. 43. 44. 45. 46. 61 monolayer permeability and modulation of cell surface coagulant properties. J. Exp. Med. 170, 1387–1407. , and Bucala, R. (1992) Immunochemical detection of advanced glycosylation end products in vivo. J. Biol. Chem. 267, 5133–5138. , et al. (1992) Isolation and characterization of two binding proteins for advanced glycosylation endproducts from bovine lung which are present on the endothelial cell surface. J. Biol. Chem. 267, 14987–14997. Westwood, M. E. and Thornalley, P.
Laser confocal unit (Yokogawa Electric Corp, CSU10, Tokyo, Japan). a. Confocal light microscope (Olympus Tokyo, Japan). b. 3CCD Camera (Hamamtsu Photonics, Hamamatsu, Japan). c. Monitor. From: Methods in Molecular Biology, vol. 196: Oxidants and Antioxidants: Ultrastructure and Molecular Biology Protocols Edited by: D. , Totowa, NJ 31 32 Majima et al. d. Laser system (Yokogawa). e. Macintosh computer. 2. Reagents and Supplies 1. 0 poly-d-lysine coated, MatTek Corporation, MA). 2. 4% buffered paraformaldehyde.
The two-step procedure for the histochemical detection of protein carbonyl groups (oxidized proteins). 2. Reagents 1. 2. 3. 4. 5. 6. 7. 8. 4. 2,4-dinitrophenyl-hydrazine (2,4-DNPH) (Merck, Cat. # 103081) (see Note 1). 3-amino-9-ethylcarbazole (AEC) (Sigma, Cat. # A6926) (see Note 2). T. Compound, Electron Microscopy Science, Cat. # 62550-01). Serial dilutions of ethanol: 100, 95, 70, 50, 30% in distilled water. Rabbit anti-dinitrophenyl IgG (Dako Cat. # V0401). Peroxidase-labeled sheep anti-rabbit IgG (Roche Diagnostic, Cat.