![]() Panel f represents control cells with no primary antibody to assess background. Panel e represents serum starved cells with reduced signal. ![]() Panel d represents the merged image showing nuclear localization. F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). The cells were labeled with Ki-67 Monoclonal Antibody (SolA15), eBioscience™ (Product # 14-5698-82) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A1-1006, 1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). ![]() The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. Immunofluorescence analysis of Ki67 was performed using 70% confluent log phase HeLa cells serum starved for 36 Hrs followed by serum release for 6 Hrs. ![]()
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