Anti kat6b antibody

Isolated chondrocytes were collected for protein extraction using RIPA lysis buffer. A BCA Protein Concentration Assay Kit (Thermo Scientific, USA) was used to determine the protein concentration of the sample according to the manufacturer’s protocol. Proteins were separated on a 10% SDS gel and transferred into PVDF membranes. Primary antibodies were used as follows: anti-KAT6B antibody, 1:1000 dilution, cat no. ab191994, Abcam; anti-RUNX2 antibody, 1: 1000 dilution, cat no. ab76956, Abcam; anti-RUNX3 antibody, 1: 1000 dilution, cat no. ab49117, Abcam; anti-COL2A1 antibody, 1: 100 dilution, cat no. ab239007, Abcam; anti-COL10A1 antibody, 1: 1000 dilution, cat no. ab182563, Abcam; anti-β-catenin antibody, 1:1000 dilution, cat no. ab6302, Abcam; anti-c-Myc antibody, 1:1000 dilution, cat no. ab32072, Abcam; anti-VEGF antibody, 1:1000 dilution, cat no. AB-293-NA, Bio-Techne; anti-GAPDH antibody, 1:3000 dilution, cat no. sc-47724, Santa Cruz Biotechnology. After primary antibody incubation, the membranes were incubated with a secondary anti-rabbit antibody or anti-mouse antibody (1: 5000, Cell Signaling Technology, USA). The bands were measured using a chemiluminescence system (Bio-Rad, USA) imaging system.

The tissues were cut into 4 μm slides. After blocking with nonfat 5% milk for 60 min at 37 °C, the slides were located with primary antibodies (anti-KAT6B antibody, 1: 200 dilution, cat no. ab191994, Abcam) at 4 °C overnight. The slides were found with appropriate secondary antibodies at 37 °C for 1 h and co-stained with hematoxylin for 2 min at room temperature. The images were acquired using a microscope (Nikon ECLIPSE 80i, Nikon Corporation, Tokyo, Japan).