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
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Fig1:
Western blot analysis of EEA1 on different lysates with Rabbit anti-EEA1 antibody (R1401-23) at 1/1,000 dilution. Lane 1: HAP1-parental cell lysate Lane 2: HAP1-EEA1 KD cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 162 kDa Observed band size: 162 kDa Exposure time: 40 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (R1401-23) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
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 |
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Fig3: 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. |
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Fig4: 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. |
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Fig5: 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. |
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Fig6: 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”.