VALIDATION IMAGES
4% Paraformaldehyde-fixed Human Umbilical Artery Endothelial Cells(HUAEC)probed with Anti-Vimentin Polyclonal Antibody, Alexa Fluor 488 conjugated (bs-8533R-A488) 1:100, 60 minutes at 37°C. DAPI(5ug/ml,blue) was used to stain the cell nuclei
Lymph node lysates probed with Vimentin Polyclonal Antibody, Unconjugated (bs-8533R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
Lane 1: Hela KO Vimentin cell lysates; Lane 2: Hela cell lysates probed with Vimentin Polyclonal Antibody, Unconjugated (bs-8533R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: U-87MG cell lysates; Lane 2: Mouse NIH/3T3 cell lysates probed with Vimentin Polyclonal Antibody, Unconjugated (bs-8533R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Tissue/cell: U-87MG cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Vimentin) Polyclonal Antibody, Unconjugated (bs-8533R)antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
A549 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Vimentin Polyclonal Antibody(bs-8533R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Lane 1: Human A549 cell lysates; Lane 2: Human HUVEC cell lysates; Lane 3: Human U251 cell lysates probed with Vimentin Polyclonal Antibody, Unconjugated (bs-8533R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.