Hmox1 antibody

For confirming hub genes’ diagnostic value, we first normalized the gene expression data of GSE58438, GSE71647, and GSE58438 and synthesized them into a combined dataset using the min-max normalization method. ROC curves were then examined to determine accuracy, and hub genes with an AUC greater than 0.7 were considered diagnostic. p < 0.05 reported statistical significance. In addition, we used WB with the Hmox1 antibody from Proteintech (10701-1-AP) to determine hub gene expression in the renal IRI and sham groups. Kidney tissues from each group underwent 15 min of homogenization and centrifugation at 4 °C, 12,000 rpm, followed by collection of the supernatant. The bicinchoninic acid assay determined the protein concentration. Then, corresponding tissue proteins underwent sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and were transferred onto a membrane, which was incubated in a primary antibody solution (1:1000 dilution) at 4 °C overnight. On the following day, the strips were washed with Tris-buffered saline containing 0.1% Tween-20 detergent, underwent 1 h of incubation of secondary antibody at room temperature (20 °C), and were washed again. The bands were assessed using chemiluminescence, and the figures were analyzed using ImageJ software. Statistical significance was set at p < 0.05 to determine the difference between groups using Prism 9.