Flow cytometric analysis of ERK1 expression in C2C12 cells using ERK1 antibody (R021584, 1:2,000). Green, isotype control; red, ERK1.

Validation of ERK1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with ERK1 antibody (R021584, 1:2,000) and analyzed using BD flow cytometer.

Immunocytochemical staining of HepG2 cells with ERK1 antibody (R021584, 1:1,000). Nuclei were stained blue with DAPI; ERK1 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 sigmoid colon carcinoma using ERK1 antibody (R021584, 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 ERK1 antibody (R021584). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with ERK1 antibody (R021584, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using ERK1 antibody (R021584). ERK1 expression in wild type (WT) and ERK1 shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with ERK1 antibody (R021584, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.