Rabbit anti phospho nf kb p65 ser536

The collected protein samples were quantified using BCA method (Yeasen, China) and separated by 10% SDS-PAGE. Then, the protein was transferred onto the PVDF membrane (Millipore, Germany). After blocking with 5% non-fat milk at room temperature for 1 h, the membrane was incubated with primary antibodies including rabbit anti-PER1 (Affinity, USA, 1:800), rabbit anti-MMP13 (Proteintech, China, 1:1000), rabbit anti-NF-kB p65 (Affinity, USA, 1:500), rabbit anti-Phospho-NF-kB p65 (Ser536) (Affinity, USA, 1:500) and rabbit anti-GAPDH (Affinity, USA, 1:2000) at 4 °C for 16 h. Following the washing with 0.1% TBST, the secondary antibody (Beyotime, China) was incubated with the membrane. After washing with 0.1% TBST, a chemiluminescence ECL system (Millipore, Germany) was used to visualize the immunoreactive proteins.

The collected protein samples were quantified using BCA method (Yeasen, China) and separated by 10% SDS-PAGE. Then, the protein was transferred onto the PVDF membrane (Millipore, Germany). After blocking with 5% non-fat milk at room temperature for 1 h, the membrane was incubated with primary antibodies including rabbit anti-PER1 (Affinity, USA, 1:800), rabbit anti-MMP13 (Proteintech, China, 1:1000), rabbit anti-NF-kB p65 (Affinity, USA, 1:500), rabbit anti-Phospho-NF-kB p65 (Ser536) (Affinity, USA, 1:500) and rabbit anti-GAPDH (Affinity, USA, 1:2000) at 4 °C for 16 h. Following the washing with 0.1% TBST, the secondary antibody (Beyotime, China) was incubated with the membrane. After washing with 0.1% TBST, a chemiluminescence ECL system (Millipore, Germany) was used to visualize the immunoreactive proteins.