Figure@ Identification of the EID1 protein by the monoclonal antibody clone #26 by Western blotting.
Crude cell extracts of MCF7 cells (breast cancer cell line) transfected with control vector pCMV1 (lane 1) or the EID1expression vector cDNA3/EID1 (lane 2) were analyzed by Western blotting using anti-EID1 antibody clone #26 as the primary antibody and HRP-conjugated-mouse IgG as the
secondary antibody. The EID1 protein was identified as a 21kDa protein band as shown by an arrow.
Fig. 2. Indirect immunofluorescence staining of EID1 protein by anti-EID1 antibody (clone 26)
in HeLa cell.
Hela cells grown
Fixation of the cells in 4% paraformaldehyde over night
Permeabilization in 0.25% Triton X-100/PBS for 10 min
Blocking in 1.5% BSA/PBS for 30 min
Incubation over night in anti-EID1 (clone 26) antibody diluted 1/1,000 by blocking buffer
Incubation for 60min in 2nd antibody, Goat anti-rabbit IgG conjugated with Alex 488 (1:1000 dilution)
Nuclei were stained with DAPI
The images show that majority of EID1 detected by this antibody is localized in nucleus and subfraction may be localized in cytoplasm, and that it may shuttle between them as described in some literatures.