Rabbit anti bcl 2 antibody

Total RNA was isolated using Trizol reagent and cDNAs were synthesized by a RT-PCR Kit (Takara, Japan). Specific primers were designed for RT-PCR and qPCR reaction (Table 1). The conditions and procedures were referred to the instructions. The proteins were extracted with RIPA buffer (Boster, China) and the BCA Protein Assay Kit (Boster, China) was used to detect protein concentration referring to the instructions. Proteins were isolated by SDS-PAGE and transferred to PVDF membrane. The membrane was blocked with 5% skim milk powder and then incubated with rabbit anti-ITPKB antibody, 1:200 ((Bioss, China), rabbit anti-Caspase-3 antibody, 1:300 (Bioss, China), rabbit anti-Bcl-2 antibody, 1:300 (Bioss, China) and mouse anti-β-actin antibody, 1:6000 (Bioss, China). The proteins bandings were detected with ECL Western Blotting Substrate (Invitrogen, USA).

ε-Caprolactone (ε-CL, 99%, Sigma-Aldrich) was stirred over CaH₂ for 24 h at room temperature and then it was distilled at reduced pressure under nitrogen. α-Methoxy-poly(ethylene glycol)₄₄-ω-hydroxide (MeO–PEG₄₄–OH, Mn = 2000 g/mol) was dried by co-evaporation with anhydrous toluene using a rotary evaporator. Methane sulfonic acid (MSA) was purchased from Sigma-Aldrich, and was used without further purification. Rabbit α-tubulin antibody and mouse β-Actin were purchased from Sigma-Aldrich; rabbit anti-CD31 (GB11063-2) was purchased from Servicebio; rabbit anti-Bcl-2 antibody and rabbit anti-Ki67 were purchased from Bioss; rabbit anti-MDR1/ABCB1 (E1Y7B) antibody was purchased from Cell Signaling Technology; horseradish peroxidase (HRP)-conjugated secondary antibodies were obtained from Cell signaling Technology. LysoTracker^® Red DND-99 and DAPI were obtained from Thermo Scientific. Reactive oxygen species (ROS) detection kit and Annexin V-FITC apoptosis detection kit were purchased from Biovision. ECL western blotting detection kit was purchased from Millipore.