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2 protocols using rabbit anti mouse igg igm h l hrp secondary antibody

1

Cell Viability, Apoptosis, and Oxidative Stress Assays

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RPMI 1640 medium, RPMI red-phenol-free medium, fetal bovine serum (FBS), L-glutamine, and penicillin–streptomycin were purchased from Gibco BRL (Cergy-Pontoise, France). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT), human anti-β-actin antibody, cell death detection enzyme-linked immunosorbent assayPLUS (ELISA), and 2′,7′-dichlorofluorescéine diacetate (DCFDA) were obtained from Sigma-Aldrich (Saint-Quentin-Fallavier, France). LysoTracker, goat anti-rabbit IgG H&L horseradish peroxidase (HRP) secondary antibody, Poly-ADP-ribose polymerase (PARP) antibody, caspase-3 antibody, and cleaved caspase-3 antibody were purchased from Cell Signaling Technology—Ozyme (Saint-Quentin-en-Yvelines, France). MitoTracker, ER-Tracker, and rabbit anti-mouse IgG-IgM H&L HRP secondary antibody were obtained from Invitrogen—Thermo Fisher Scientific (Villebon-sur-Yvette, France). Immobilon Western Chemiluminescent HRP Substrate was acquired from Merck (Lyon, France).
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2

Autophagy Modulation in Cellular Studies

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RPMI-1640 medium, fetal bovine serum (FBS), L-glutamine and penicillin-streptomycin were purchased from Gibco BRL. Diosgenin [(25R)-5-spirosten-3β-ol], 3-methyladenine (3-MA), metformin (Met) were obtained from Sigma-Aldrich. The monoclonal antibody against Beclin1, Atg7, Atg12-5, Atg3, LC3A/B were acquired from Cell Signaling Technology-Ozyme (Saint-Cyr-L′école, France). GAPDH antibody was purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology-Clinisciences (Nanterre, France). Goat anti-rabbit IgG H&L horseradish peroxidase (HRP) secondary antibody was purchased from Abcam (Paris, France). Rabbit anti-mouse IgG-IgM H&L HRP secondary antibody and propidium iodide (PI) were obtained from Invitrogen-Thermo Fisher Scientific, Inc. Immobilon Western Chemiluminescent HRP Substrate was acquired from Merck.
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