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2 protocols using nbt chloride tablets

1

Dihydrolipoamide Synthesis and Antibody Generation

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Dihydrolipoamide was synthesized from lipoamide using sodium borohydride as previously described 34 (link), 35 . All PCR primers were purchased from Life Technologies (Carlsbad, CA). ε-amino-N-caproic acid was obtained from MP Biochemicals. Acrylamide/bisacrylamide, ammonium persulfate, Bradford protein assay solution, and Coomassie brilliant blue (CBB) R-250 were from Bio-Rad laboratories (Richmond, CA, USA). NADH, BSA, lipoamide, EDTA, and NBT chloride tablets were obtained from Sigma (St. Louis, MO, USA). Serva Blue G was purchased from Serva (Heidelberg, Germany). Rabbit anti-DLDH polyclonal antibodies (IgG) and goat anti-rabbit IgG conjugated with horseradish peroxidase were purchased from US Biological (Swampscott, MA, USA) and Invitrogen (San Diego, CA, USA), respectively. Hybond-C membrane and an ECL immunochemical detection kit were obtained from GE Healthcare (Piscataway, NJ, USA).
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2

Protein Carbonyl Assay Protocol

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Biotin-linked aldehyde reactive probe ARP) for protein carbonyl assay was from Cayman Chemical (Ann Arbor, MI). Dihydrolipoamide was synthesized from lipoamide in our own laboratory using sodium borohydride as previously described [29] , [30] . ε-amino-N-caproic acid was obtained from MP Biochemicals. Acrylamide/bisacrylamide, ammonium persulfate, Bradford protein assay solution, coomassie brilliant blue (CBB) R-250, immunoblotting membrane, and an ECL immunochemical detection kit were from Bio-Rad laboratories (Richmond, CA, USA). NADH, BSA, lipoamide, EDTA, ATP, and NBT chloride tablets were obtained from Sigma (St. Louis, MO, USA). Serva Blue G was purchased from Serva (Heidelberg, Germany). Anti-PARP antibody was purchased from Trevigen (Gaithers burg, MD). Anti-NQO1 antibodies were from Sigma. Rabbit anti-DLDH polyclonal antibodies (IgG) and goat anti-rabbit IgG conjugated with horseradish peroxidase were purchased from US Biological (Swampscott, MA, USA) and Invitrogen (San Diego, CA, USA), respectively. Other antibodies were from Abcam (Cambridge, MA).
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