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2 protocols using catalase d4p7b

1

Evaluating Oxidative Stress Response

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Lomefloxacin hydrochloride, penicillin G, amphotericin B, H2DCFDA (2′,7′-dichlorofluorescein diacetate), SIGMAFAST™ Protease Inhibitor Cocktail Tablet, and Phosphatase Inhibitor Cocktail 3, Dulbecco’s phosphate-buffered saline (DPBS) with MgCl2 and CaCl2, phosphate buffered saline (PBS),and Fibroblast Growth Medium were obtained from Sigma Aldrich Inc. (St. Louis, MO, USA). A Pierce BCA Protein Assay Kit, ECL Western Blotting Substrate, and Hoechst 33342, CellROX™ Green Reagent were obtained from Thermo Fisher Scientific (Waltham, MA, USA). GAPDH (14C10) Rabbit mAb, SOD1 (71G8) Mouse mAb, SOD2 (D9V9C) Rabbit mAb, Catalase (D4P7B) Rabbit mAb, and GPx1 (C8C4) Rabbit mAb were obtained from Cell Signaling (Danvers, MA, USA), and Anti-Rabbit IgG (A154), Anti-Mouse IgG, Tween-20, RIPABuffer and PVDF membranes were obtained from Sigma-Aldrich Inc. (St. Louis, MO, USA). Neomycin sulfate was obtained from Amara (Kraków, Poland). Trypsin/EDTA solution was purchased from Cascade Biologics/Gibco (Carlsbad, CA, USA). Solution 3 (1 μg/ mL DAPI, 0.1% triton X-100 in PBS), Solution 5 (VB-48TM, propidium iodide-PI, acridine orange—AO), NC-Slide A8 and Via-1-Cassette (AO and DAPI fluorophores) were obtained from ChemoMetec (Lillerød, Denmark). Cell Proliferation Reagent WST-1 was produced by Roche GmbH (Mannheim, Germany). Other chemicals were from POCH S.A. (Gliwice, Poland).
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2

Antibody Characterization for IF and WB

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Antibodies used for immunofluorescence and Western blotting, included: NQO1 (A180), PARP1 (SC-8007, Santa Cruz, La Jolla, CA), Actin (C4, Santa Cruz), PAR (Trevigen, Gaithersburg, MD), Cleaved caspase 7 (D6H1, Cell Signaling, Danvers, MA), cleaved caspase 3 (5A1E, Cell Signaling), p53 (DO-1, Santa Cruz), catalase (D4P7B, Cell Signaling), γH2AX (JBW301, Millipore, Temecula, CA), and α-tubulin (Santa Cruz).
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