EEA1 Rabbit Polyclonal Antibody
cat.: R1401-23
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: 162 kDa
Isotype: IgG
Immunogen: Synthetic peptide within N-terminal human EEA1.
Positive control: NIH/3T3, Hela, Jurkat, 293T, A431, rat brain tissue, rat prostate tissue, mouse colon tissue.
Subcellular location: Cytoplasm, early endosome membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC

1:1,000
1:100-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: Q15075 Human | Q8BL66 Mouse
Alternative names: Early endosome antigen 1 Early endosome antigen 1, 162kD Early endosome associated protein EEA 1 EEA1 EEA1_HUMAN Endosome associated protein p162 Endosome-associated protein p162 MST105 MSTP105 ZFYVE2 Zinc finger FYVE domain containing protein 2 Zinc finger FYVE domain-containing protein 2
Images
R1401-23_1.jpg Fig1: Western blot analysis of EEA1 on different lysates. 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 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: NIH/3T3 cell lysate
Lane 2: Hela cell lysate
Lane 3: Jurkat cell lysate
Lane 4: 293T cell lysate
Lane 5: A431 cell lysate
Lane 6: Rat brain tissue lysate
R1401-23_2.jpg Fig2: ICC staining EEA1 in NIH/3T3 cells (green). 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 the antibody (R1401-23) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution.
R1401-23_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded rat prostate 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 (R1401-23) at 1/50 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.
R1401-23_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse colon 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 (R1401-23) at 1/50 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.
R1401-23_5.jpg Fig5: Flow cytometric analysis of EEA1 was done on Hela cells. The cells were fixed, permeabilized and stained with EEA1 antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.