Rabbit monoclonal anti histone h3 d1h2

Palmitic acid (PA, sodium salt) and cis-4,7,10,13,16,19-docosahexaenoic acid (DHA, sodium salt) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Palmitic acid was dissolved in 50% ethanol, while docosahexaenoic acid was dissolved in 100% ethanol to a concentration of 10 mg/mL; these stock solutions were stored at −80 °C under N₂ until use. The rabbit monoclonal anti-Calnexin (C5C9) and the rabbit monoclonal anti-Histone H3 (D1H2) antibodies were purchased from Cell Signaling Technology, Danvers, MA, USA. The mouse monoclonal anti-HSP60 (66041) antibody was purchased from Proteintech, Deansgate, Manchester, UK. Bound primary antibody was visualized by proper secondary horseradish peroxidase (HRP)-linked antibody, purchased from Cell Signaling Technology, Danvers, MA, USA. All solvents were purchased from Carlo Erba Reagents, Italy. Avanti Polar Lipids Inc. (Alabaster, AL, USA) supplied the fatty acid methyl ester (FAME) standards for GC analysis and neutral lipids (TG and CE) for LC analysis. Phospholipid standards were purchased from Sigma Aldrich, St. Louis, MO, USA.

The cytoplasmic, membrane and organelle and cytoskeletal and nuclear fractions of cells were obtained by lysing using the Cell Fractionation Kit (#9038, Cell Signaling Technology). Whole‐cell proteins were extracted by lysing in RIPA buffer. The protein lysate was separated using 10% sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and electro‐transferred onto a polyvinylidene fluoride membrane. The membranes were blocked in 5% BSA and were incubated with primary antibodies overnight at 4°C. The following antibodies were used: mouse monoclonal anti‐NUMB (1:1000, ab14140; Abcam), mouse monoclonal anti‐MDM2 [2A10] (1:50, ab16895; Abcam, Cambridge, MA, USA), Rabbit Polyclonal anti‐P53 (1:1000, #9282; Cell Signaling Technology), Rabbit Monoclonal anti‐Histone H3 (D1H2) (1:2000, #4499; Cell Signaling Technology), Rabbit Polyclonal anti‐Caveolin‐1 (1:1000, bs‐1453R; Bioss) and mouse anti‐GAPDH (1:1000, 60 004‐1‐Ig; Proteintech Group). The membranes were washed and incubated with anti‐rabbit or anti‐mouse secondary antibody (1:5000, SSA004/SSA007; Sino Biological Inc) for 2 hours at room temperature. Finally, antigen‐antibody complexes were detected using an electrochemiluminescence Western blotting detection reagent.