Rabbit anti ppar α antibody

Immunohistochemical analysis was performed using the Histone Simple Stain Kit (Nichirei, Tokyo, Japan), according to the manufacturer's instructions. Paraffin-embedded sections were deparaffinised with xylene and then rehydrated in a descending series of ethanol washes. The sections were treated for 15 minutes with 3% H₂O₂ in methanol to inactivate endogenous peroxidases and then incubated at room temperature for 1 hour with primary antibodies to PPARα (rabbit anti-PPARα antibody, 1:200; Proteintech, Wuhan, China) and P53 (rabbit anti-P53 antibody, 1:200; Proteintech). Tissue sections were observed with a microscope (Olympus, Tokyo, Japan).

Proteins were extracted from kidney tissue using radio immunoprecipitation assay buffer (P0013B; Beyotime, Shanghai, China). Samples were electrophoresed on a 10% SDS-PAGE gel, and proteins were transferred to polyvinylidene fluoride membrane (Immobilon, Millipore, Billerica, MA, USA). Membranes were blocked in Tris-buffered saline with 0.1% Tween-20 (TBS-T) containing 5% skim milk and then incubated in primary antibody diluent (P0023A; Beyotime) and gently shaken overnight at 4 °C. Primary antibodies against PPARα (rabbit anti-PPARα antibody, 1:1000; Proteintech), P53 (rabbit anti-P53 antibody, 1:1000; Proteintech), and anti-β-actin (1:1000; Cell Signaling Technology) were utilised. Membranes were then incubated with secondary antibody (anti-rabbit Ig-G, 1:1000; Cell Signaling Technology for 1 hour). This analysis was carried out independently three times. Protein levels are expressed as protein/β-actin ratios to minimise loading differences. The relative signal intensity was quantified using NIH ImageJ software.