Western blotting analysis using vimentin antibody (Cat#3238). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with vimentin antibody (Cat#3238, 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 endometrial carcinoma using vimentin antibody (Cat#3238, 1:100). 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.
Immunohistochemistry analysis was performed on mouse brain sections (ARH, arcuate nucleus of the hypothalamus) fixed in 10% formalin and permeabilized with 0.1% Triton X-100. Sections were incubated overnight at 4°C with vimentin antibody (Cat#3238, 1:1,000), followed by incubation with Goat Anti-Rabbit Alexa Fluor® Cy3 secondary antibody (1:500) for 2 hours at room temperature (red). Cell nuclei are indicated by DAPI (blue).
Image courtesy of Yong Xu, MD, PhD from Morsani College of Medicine, University of South Florida (Tampa Campus).
Immunocytochemical staining of HepG2 cells with Vimentin antibody(Cat#3238, 1:1,000). Nuclei were stained blue with DAPI; Vimentin was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: High. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Scale bar: 20 μm.
Flow cytometric analysis of vimentin expression in HepG2 cells using vimentin antibody (Cat#3238, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use. Green, isotype control; red, vimentin.
