Flow cytometric analysis of Smad4 expression in C2C12 cells using Smad4 antibody (R020324,1:2,000). Green, isotype control; red, Smad4.

Immunocytochemical staining of C2C12 cells with Smad4 antibody (R020324, 1:1,000). Nuclei were stained blue with DAPI; Smad4 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar: 20 μm.

Immunohistochemistry was performed on paraffin-embedded human breast carcinoma using smad4 antibody (R020324, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar: 25 μm.

Western blotting analysis using Smad4 antibody (R020324). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Smad4 antibody (R020324,1:5,000) and HRP-conjugated goat anti rabbit secondary antibody (1:50,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using Smad4 antibody (R020324). Smad4 expression in wild type (WT) and Smad4 shRNA knockdown (KD) HT-1080 cells with 30 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with Smad4 antibody (R020324,1:5,000) and HRP-conjugated goat anti rabbit secondary antibody (1:50,000) respectively. Image was developed using ECL Substrate Kit.