Tak1 antibody

The kidney tissue sections were deparaffinized and antigen retrieval was performed using a microwave oven at high temperature (300 W) for 30 min. Following the exhaustion of endogenous peroxidase with methanol and hydrogen peroxide for 30 min at room temperature, the slides were blocked with 0.5% bovine serum albumin (Sigma-Aldrich; Merck KGaA) for 60 min at 37°C and incubated with TAK1 antibody (1:200 dilution; cat. no. ab109526; Abcam) overnight at 4°C. The SABC staining kit (cat. no. SA1026; Boster Biological Technology) was used to perform the chromogenic reaction. Following rinsing in PBS, the samples were incubated with the mouse anti-rabbit antibody (cat. no. SA1026; Boster Biological Technology) at 37°C for 30 min, followed by rinsing in PBS three times for 5 min. The samples were covered in a drop of DAB developing solution, cell nuclei were stained with hematoxylin at room temperature for 5 min, following dehydration with an alcohol gradient, the samples were cleared with dimethylbenzene xylene and mounted with neutral gum sealant. Photomicrographs of different fields of view were captured using an Olympus DSX100 optical microscope (Olympus Corporation; magnification, ×400). and the average optical density (OD) was calculated using Image-pro plus 6.0 analysis software (Media Cybernetics, Inc.).