EEA1-Early Endosome Marker Mouse monoclonal Antibody IgG1
Fig1: Western blot analysis of EEA1 on 293 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:2,000 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Fig2: ICC staining EEA1 in 293 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with EEA1 monoclonal antibody at a dilution of 1:200 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™555 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution.
Fig3: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-EEA1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (M1503-1) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
Host Species; Species ReactivityMouse; Human, Mouse, Rat
ImmunogenSynthetic Peptide of the N terminal residues of Human EEA1.
Application SummaryWB, ICC, IHC
Purification; FormulationProA affinity purified; 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide.; Liquid form.
ALTnamesEarly endosome antigen 1,Endosome-associated protein p162,Zinc finger FYVE domain-containing protein 2
BackgroundEEA1 localizes exclusively to early endosomes and has an important role in endosomal trafficking. EEA1 binds directly to the phospholipid phosphatidylinositol 3-phosphate through its C-terminal FYVE domain and forms a homodimer through a coil. EEA1 acts as a tethering molecule that couples vesicle docking with SNAREs such as N-ethylmaleimide sensitive fusion protein, bringing the endosomes physically closer and ultimately resulting in the fusion and delivery of endosomal cargo. Due to the proteins importance in vesicular trafficking, a number of intracellular bacteria prevent EEA1 recruitment to the vacuole.Mycobacterium tuberculosis is known to inhibit the recruitment of EEA1 to the phagosomal membrane through CamKII. Legionella pneumophila also prevents EEA1 recruitment through a currently unknown mechanism. Interestingly, the related pathogen Legionella longbeachae recruits EEA1 and appears to replicate within a modified early endosome.(M1503-1)