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Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the <t>SYPRO</t> <t>Ruby-stained</t> two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).
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Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the <t>SYPRO</t> <t>Ruby-stained</t> two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).
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Bio-Rad sypro ruby 146 fluorescence dye
Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the <t>SYPRO</t> <t>Ruby-stained</t> two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).
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Thermo Fisher sypro ruby protein gel stain
Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the <t>SYPRO</t> <t>Ruby-stained</t> two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).
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Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the <t>SYPRO</t> <t>Ruby-stained</t> two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).
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Bio-Rad sypro ruby stained sds page
Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the <t>SYPRO</t> <t>Ruby-stained</t> two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).
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Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the <t>SYPRO</t> <t>Ruby-stained</t> two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).
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Bio-Rad molecular imager fx pro plus
Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the <t>SYPRO</t> <t>Ruby-stained</t> two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).
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Image Search Results


Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the SYPRO Ruby-stained two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).

Journal: The Journal of Biological Chemistry

Article Title: Mitochondrial Alterations and Oxidative Stress in an Acute Transient Mouse Model of Muscle Degeneration

doi: 10.1074/jbc.M113.493270

Figure Lengend Snippet: Proteomic identification of oxidized proteins in DMD. A, total extracts of DMD (n = 6) and control (n = 6) were derivatized with DNPH and subjected to two-dimensional PAGE followed by anti-DNP Western blot. The protein spots on the Western blot were compared with the SYPRO Ruby-stained two-dimensional gel of total protein extract from DMD muscle to identify oxidized proteins common between control and DMD (outlined by a complete circle) and those specific to DMD (outlined by broken circle). There were 23 distinct spots in the DMD blot, of which 8 (numbered 16–23) were not represented in control. Seven spots (numbers 17 to 23) were excised out for analysis by mass spectrometry. B, list of the spot numbers and the corresponding muscle proteins that are potentially oxidized in DMD patients compared with control. C, the activity of aconitase, one of the proteins carbonylated in the DMD muscle is significantly decreased. Activity indicated in % (100% activity = 0.069 mm/min/mg protein); *, p < 0.05 compared with control (C). D, aconitase activity (in mouse muscle) is inhibited by H2O2. **, p < 0.01 compared with untreated control (C).

Article Snippet: IPG strips, Coomassie Brilliant Blue G-250, and SYPRO Ruby were obtained from Bio-Rad Laboratories.

Techniques: Control, Western Blot, Staining, Two-Dimensional Gel Electrophoresis, Mass Spectrometry, Activity Assay