Ab133691

Lysis buffer was determined by radioimmunoprecipitation and protein concentration by the bovine serum albumin method. Proteins were isolated from the lysate by SDS-PAGE and transferred to a PVDF membrane. After containment, specific primary resistance to PKM2 (1/1000; ab85555; Abcam, USA), HK2 (1/1000; Ab133691), LDHA (1/5000; Ab52488), CD36 (1/1000; Ab133625), TSG101 (1/1000; Ab133586), PARP (1/1000; Ab32071), Bax (1/1000; Ab32503), bcl-2 (1/2000; Ab182858) or beta-actin (1/200; Ab115777) for incubation followed by peroxidase-coupled secondary resistance for incubation. Final detection of protein traces using an improved chemiluminescence detection system (Pierce, USA).

Tissues or cells were added with RIPA lysate to extract total cell protein. Protein concentration was determined by BCA method. The 30 μg protein sample was mixed with the loading buffer thoroughly. The protein was isolated by sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE). After the reaction, the membrane was transferred and sealed, and the primary protein antibody including RCN1 (1/1000; ab205927; Abcam, USA), DNAJA2 (1/10000; ab168365; Abcam, USA), LASP1 (1/10000; ab156872, USA), IBSP (1/1000; A-AP14114a; Abgent, USA), β-actin (1/5000; ab179467; Abcam, USA), LAT2 (1/1000; ab75610; Abcam, USA), FUZ (1/1000; ab111842; Abcam, USA), HOOK2 (1/1000; ab133691; Abcam, USA), and DAPK2 (1/1000; ab111928; Abcam, USA) was added and incubated at 4°C for 24 h. After TBST washing, IgG secondary antibody (1/5000; ab7090; Abcam, USA) was added. ECL was added, and the protein bands were analyzed by automatic gel imaging system.