Immunocytochemistry analysis of K-Ras in HeLa cells. Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were incubated with K-Ras monoclonal antibody (Product # MA5-50601) with a dilution of 1:200 in 2% BSA overnight at 4 ℃. Followed by secondary antibody Goat Anti-Mouse IgG H&L (iFluor™ 488) at a dilution of 1:1,000. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Western blot analysis of K-Ras in different various lysates. Lane 1: Rat brain tissue lysate (20 µg/Lane); Lane 2: Jurkat cell lysate (10 µg/Lane); Lane 3: N2A cell lysate (10 µg/Lane). Predicted band size: 21 kDa. Observed band size: 21 kDa. Exposure time: 2 minutes; 15% SDS-PAGE gel. Primary antibody K-Ras monoclonal antibody (Product # MA5-50601) with a dilution of 1:500 was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody with a dilution of 1:100,000 was used for 1 hour at room temperature. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature.
Supplier Page from Thermo Fisher Scientific for K-Ras Antibody