PRODUCT CODE: R1401-23

EEA1 Rabbit Polyclonal Antibody (R1401-23)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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.<br />
Positive control: <br />
Lane 1: NIH/3T3 cell lysate<br />
Lane 2: Hela cell lysate<br />
Lane 3: Jurkat cell lysate<br />
Lane 4: 293T cell lysate<br />
Lane 5: A431 cell lysate<br />
Lane 6: Rat brain tissue lysate
  • 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.<br />
Positive control: <br />
Lane 1: NIH/3T3 cell lysate<br />
Lane 2: Hela cell lysate<br />
Lane 3: Jurkat cell lysate<br />
Lane 4: 293T cell lysate<br />
Lane 5: A431 cell lysate<br />
Lane 6: Rat brain tissue lysate
  • 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.
  • 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.
  • 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.
  • 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.
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

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

EEA1 Rabbit Polyclonal Antibody (R1401-23)

Immunogen

Synthetic peptide of the n terminal residues of human eea1.

Host

Rabbit

Positive Control

NIH/3T3, Hela, Jurkat, 293T, A431, rat brain tissue, rat prostate tissue, mouse colon tissue.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Peptide affinity purified.

MOLECULAR WEIGHT

162 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB:1:1,000

  • ICC:1:100-1:200

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

EEA1

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; ZFYVE2 antibody; Zinc finger FYVE domain containing protein 2 antibody; Zinc finger FYVE domain-containing protein 2 antibody

SUBCELLULAR LOCATION

Cytoplasm, early endosome membrane.

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.