Description
Vimentin (VIM) is a type III intermediate filament (IF) protein that is expressed inmesenchymal cells. Vimentin is the major cytoskeletal component ofmesenchymal cells. Because of this, vimentin is often used as a marker ofmesenchymally-derived cells or cells undergoing an epithelial-to-mesenchymaltransition (EMT) during both normal development and metastatic progression. Besides, Keratin 20 (KRT20) has been identified as an interactor of VIM, thus abinding ELISA assay was conducted to detect the interaction of recombinanthuman VIM and recombinant human KRT20. Briefly, VIM were diluted serially inPBS, with 0.01% BSA (pH7.4). Duplicate samples of 100ul then transferred to KRT20-coated microtiter wells and incubated for 2h at 37 degree C. Wells were washedwith PBST and incubated for 1h with anti-VIM pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells wereaspirated and washed 3 times. With the addition of substrate solution, wells wereincubated 15-25 minutes at 37 degree C. Finally, add 50ul stop solution to the wells andread at 450nm immediately. The binding activity of VIM and KRT20 was shown in Figure 1, and this effect was in a dose dependent manner