Anti γh2ax ab26350

Cells were collected and lysed in RIPA buffer (Thermo Scientific) mixing with protease inhibitor cocktail (Sigma), the protein concentrations examination were utilizing the BCA protein assay kit (Pierce). We detected protein expression utilizing 20–60 μg of samples, samples were separated by SDS-PAGE and transferred to nitrocellulose membranes (Pall). The primary antibodies for western blotting were as follows: anti-HBP1 (11746-1-AP, Proteintech), anti-MDM2 (sc-965, Santa Cruz Biotechnology), anti-p21(K0081-3, MBL), anti-p16(10883-1-AP, Proteintech), anti-DNMT1(sc-10222, Santa Cruz Biotechnology), anti-EZH2(21800-1-AP, Proteintech), anti-FLAG (F1804, Sigma-Aldrich), anti-HA(MMS101P, Covance), anti-Muti-Ub(D058-3, MBL), anti-γ-H2AX(ab26350, Abcam), and anti-GAPDH(KM9002, Sungene). The fluorescence labeled secondary antibodies we used were as follows: anti-mouse IgG antibody DyLight 800 (610-145-121, Rockland) and anti-rabbit IgG DyLight 800 (611-145-002, Rockland). The infrared fluorescence graphic results were acquired utilizing Odyssey infrared imaging system (LI-COR Bioscience, Lincoln, NE).

Sevoflurane was purchased from Maruishi Pharmaceutical Co. (Osaka, Japan). N-acetyl-L-cysteine (NAC), 3-aminobenzamide (3AB), and hydrogen peroxide (H₂O₂) were purchased from Sigma-Aldrich Company (St. Louis, MO, United States). Lactate dehydrogenase (LDH) detection kit was from Beyotime Biotech (Nanjing, China). Nuclear and Cytoplasmic Protein Extraction Kit (P0028) and BCA Protein Assay Kit (P0010) were purchased from Beyotime Biotech (Shanghai, China). Anti-PAR polymer (AM80) was from Calbiochem company (Danvers, MA, United States). Anti-PARP-1 (ab32138), anti-AIF (ab32516), and anti-γH2AX (ab26350) were purchased from Abcam Company (Cambridge, MA, United States). Anti-ATM (sc-135663) and anti-p-ATM (sc-47739) were from Santa Cruz Biotechnology (Santa Cruz, CA, United States). Anti-8-OHdG (251640) was from Abbiotec (San Diego, CA, United States). PARP-1 small interfering RNA (SiRNA) was purchased from GenePharma Company (Suzhou, China). Anti-Histone-H3 (17168-1-AP) was from Proteintech Group. Anti-β-actin (AA128) was from Beyotime Biotech (Nanjing, China).

Anti–BCL-XL (BD-610209) was from BD Biosciences; anti-CTGF (NBP2-16026) and anti-COL1 (NBP1-30054, NB-600408) were from Novus Biologicals; anti-FN (ab2413), anti–α-SMA (ab5694), anti-P16 (ab189034, ab211542), and anti-γH2A.X (ab26350) were from Abcam; anti-VIM (3932), anti–cleaved caspase-3 (9664), anti-P53 (2524, 32532), anti-Ki67 (9129), anti–β-actin (3700), and anti-GAPDH (2118) were from Cell Signaling Technology; anti-P21 (AHZ0422) was from Invitrogen; and anti-FGF2 (05-118) and FGF2 neutralizing antibody (05-118) were from MilliporeSigma. Secondary antibodies for immunoblot analysis were from Jackson ImmunoResearch Laboratories. Cisplatin (P4394) and crystal violet (C0775) were from MilliporeSigma. ABT-263 (201970) was from MedKoo Biosciences. FST (528-48-3) was from Selleck Chemicals.