EEA1 Rabbit Polyclonal Antibody
Catalog# R1401-23
EEA1 Rabbit Polyclonal Antibody
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WB
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IF-Cell
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
EEA1 Rabbit Polyclonal Antibody
Antibody Type
Rabbit Polyclonal Antibody
Immunogen
Synthetic peptide within N-terminal human EEA1.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC
Target Molecular Weight
162 kDa
Positive Control
Hela, HEK-293, NIH/3T3, C6, rat prostate tissue, mouse colon tissue.
Conjugation
unconjugated
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Immunogen affinity purified.
Application Dilution
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WB
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1:1,000
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IF-Cell
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1:100-1:200
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
Target
Function
EEA1 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 coiled 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.
Background References
1. Gaullier J.-M et al. Interaction of the EEA1 FYVE finger with phosphatidylinositol 3-phosphate and early endosomes. Role of conserved residues. J Biol Chem 275:24595-24600 (2000).
2. Kauppi M et al. The small GTPase Rab22 interacts with EEA1 and controls endosomal membrane trafficking. J Cell Sci 115:899-911 (2002).
3. Merithew E et al. Determinants of Rab5 interaction with the N-terminus of early endosome antigen 1. J Biol Chem 278:8494-8500 (2003).
Subcellular Location
Cytoplasm, early endosome membrane.
Synonyms
Early endosome antigen 1 antibody
Early endosome antigen 1, 162kD antibody
Early endosome associated protein antibody
EEA 1 antibody
EEA1 antibody
EEA1_HUMAN antibody
Endosome associated protein p162 antibody
Endosome-associated protein p162 antibody
MST105 antibody
MSTP105 antibody
ExpandImages
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☑ Knockdown (KD)
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. -
Western blot analysis of EEA1 on different lysates with Rabbit anti-EEA1 antibody (R1401-23) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: C6 cell lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 25 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: R1401-23, 1/1,000 5% NFDM/TBST, overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 162 kDa
Observed band size: 162 kDa -
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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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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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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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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Alternative Products
EEA1 Recombinant Rabbit Monoclonal Antibody [JE59-34]
Application: WB,IF-Cell,FC
Reactivity: Human,Mouse,Rat,Green monkey
Conjugate: unconjugated
