Western blotting analysis using CD146 antibody (Cat#61487). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with CD146 antibody (Cat#61487, 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 CD146 antibody (Cat#61487). CD146 expression in wild type (WT) and CD146 shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with CD146 antibody (Cat#61487, 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).
Immunohistochemistry was performed on paraffin-embedded human melanoma using CD146 antibody (Cat#61487, 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 CD146 antibody (Cat#61487, 1:1,000) . Nuclei were stained blue with DAPI; CD146 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.
Flow cytometric analysis of CD146 expression in HepG2 cells using CD146 antibody (Cat#61487, 1:2,000). Green, isotype control; red, CD146.
Validation of CD146 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with CD146 antibody (Cat#61487, 1:2,000) and analyzed using BD flow cytometer.
