Western blot analysis of p57Kip2 was performed by loading 25 µg of Jurkat (lane 1), SH-SY5Y (lane 2) and Neuro-2a (lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a p57Kip2 monoclonal antibody (Product # MA5-11309) at a dilution of 1:50 (Jurkat) and 1:100 (SH-SY5Y and Neuro-2a) overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~57 kDa.