Anti gapdh antibody ab8245

We extracted total protein from articular cartilage homogenate with 1 ml of tissue lysate and protease inhibitor, and collected the supernatant after centrifugation at 12,000 rpm for 15 min at 4 °C, then determined the protein concentration via a BCA Protein Assay Kit (Beyotime Biotechnology Co.). An equal amount of protein was put into gels and resolved. Proteins were transferred to polyvinylidene difluoride membranes and exposed first to anti-MMP13 antibody ab39012, anti-TLR4 antibody ab13556, and anti -GAPDH antibody (ab8245) (Abcam, UK) and then to goat anti-rabbit IgG H&L (HRP) ab6721 (Abcam, MA). Bands were detected and analyzed using ChemiDoc™ XRS+ with Image LabTM software (BIO-RAD, USA).

Herpud1 polyclonal antibody (PA5-59485) was purchased from Invitrogen and used at a 1:2500 dilution for IF and WB. Anti-nuclear factor of activated T-cells, cytoplasmic 4 (NFATc4) antibody (ab3447) was purchased from Abcam and used at a 1:100 dilution for IF and 1:500 for WB. Anti-GAPDH antibody (ab8245) was purchased from Abcam and used at a 1:1000 dilution for WB. Anti-Lamin antibody (ab108595) was purchased from Abcam and used at a 1:1000 dilution for WB. The secondary antibodies Alexa Fluor® 488 goat anti-rabbit immunoglobulin G (IgG) (A11008) and Alexa Fluor® 633 goat anti-rabbit IgG (A21070) were purchased from Invitrogen and used at a 1:300 dilution. Alexa Fluor® 555 goat anti-rabbit IgG (ab150078) was purchased from Abcam and used at a 1:300 dilution.