Anti iκbα antibody c21

The cells were lysed with 1× RIPA buffer (diluted from 10× RIPA buffer, Cell Signaling Technology, #9806) or 1× SDS loading buffer and the supernatants were collected and applied to SDS-PAGE. Western blots were performed following the standard protocol as described (7 (link), 20 (link)). Briefly, after SDS-PAGE, the proteins were transferred to PVDF membranes (Millipore). The membranes were blocked with 5% non-fat dry milk or BSA in TBST according to primary antibody preference. Followed by primary antibody incubation, the blots were washed with TBST buffer and incubated with secondary antibodies. The antibodies included: anti-TNFAIP3 (A20) antibody (D13H3, Cell Signaling Technology, 5630S, 1:1000), anti-β-Actin antibody (C4, Santa Cruz, sc-47778, 1:1000), anti-IκBα antibody (C21, Santa Cruz, SC-371, 1:1000), NF-κB p65 Antibody (F-6, Santa Cruz, SC-8008, 1:200), anti-p-NF-κB p65 antibody (27. Ser536, Santa Cruz, SC-136548, 1:500), anti-cleaved caspase 3 antibody (Cell signaling technology, # 9661S, 1: 500). Images were captured with G: BoxXX6 (Syngene) and band intensity was determined using ImageJ, version 1.4.3.67.