Western blotting analysis using mTOR antibody (Cat#62298). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with mTOR antibody (Cat#62298, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using mTOR antibody (Cat#62298). mTOR expression in wild type (WT) and mTOR shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with mTOR antibody (Cat#62298, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunohistochemistry was performed on paraffin-embedded human breast carcinoma using mTOR antibody (Cat#62298, 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.
Immunocytochemical staining of HepG2 cells with mTOR antibody (Cat#62298, 1:1,000). Nuclei were stained blue with DAPI; mTOR was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: High. Scale bar, 20 μm.
Flow cytometric analysis of mTOR expression in HepG2 cells using mTOR antibody (Cat#62298, 1:2,000). Green, isotype control; red, mTOR.
Validation of mTOR knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with mTOR antibody (Cat#62298, 1:2,000) and analyzed using BD flow cytometer.
