Figure@Identification of the EID1 protein by the monoclonal antibody (clone #2) by Western blotteing.
Crude cell extracts of MCF7 cells (breast cancer cell line) transfected with control vector pCMV1 (lane 1) or the EID1 expression vector pcDNA3/EID1 (lane 2) were analyzed by Western blotting using anti-EID1 antibody clone #2 as the
primary antibody and HRP-conjugated-mouse IgG as the secondary antibody. The EID1 protein was identified as a 21 kDa protein band as shown by an arrow.
Fig. 2. Indirect immunofluorescence staining of EID1 protein by anti-EID1 antibody (clone 2)
in HeLa cell.. EID1 protein is localized in nuclei.
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 antibody (clone 2) diluted to 1/1,000 by blocking buffer
Incubation for 60min in 2nd antibody, Goat anti-rabbit IgG conjugated with Alex 488 (1:1,000 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.