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EEA1 Mouse Monoclonal Antibody [B0-A8]

Usd: 350

Specification

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Catalog# M1503-1

EEA1 Mouse Monoclonal Antibody [B0-A8]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

EEA1 Mouse Monoclonal Antibody [B0-A8]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide of the N terminal residues of Human EEA1.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 162 kDa

Positive Control

HeLa cell lysate, Daudi cell lysate, A431 cell lysate, SH-SY5Y cell lysate, NIH/3T3 cell lysate, C6 cell lysate, human prostate tissue, mouse prostate tissue.

Conjugation

unconjugated

Clone Number

B0-A8

RRID

AB_3073099

Product Features

Form

Liquid

Concentration

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

IgG1

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:10,000

  • IHC-P

  • 1:50-1:200

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-terminalFYVE 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. Legionella pneumophila?also prevents EEA1 recruitment through a currently unknown mechanism.[5] Interestingly, the related pathogen Legionella longbeachae?recruits EEA1 and appears to replicate within a modified early endosome.

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.

UNIPROT

SwissProt: Q15075 Human

SwissProt: Q8BL66 Mouse

SwissProt: A0A0G2K051 Rat

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

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of EEA1 on different lysates with Mouse anti-EEA1 antibody (<a href="/products/M1503-1" style="font-weight: bold;text-decoration: underline;">M1503-1</a>) at 1/10,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-EEA1 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 162 kDa<br />Observed band size: 162 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/M1503-1" style="font-weight: bold;text-decoration: underline;">M1503-1</a>) at 1/10,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of EEA1 on different lysates with Mouse anti-EEA1 antibody (M1503-1) at 1/10,000 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: 20 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 (M1503-1) at 1/10,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of EEA1 on different lysates with Mouse anti-EEA1 antibody (<a href="/products/M1503-1" style="font-weight: bold;text-decoration: underline;">M1503-1</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Daudi cell lysate<br />Lane 3: A431 cell lysate<br />Lane 4: SH-SY5Y cell lysate<br />Lane 5: NIH/3T3 cell lysate<br />Lane 6: C6 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 162 kDa<br />Observed band size: 162 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/M1503-1" style="font-weight: bold;text-decoration: underline;">M1503-1</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of EEA1 on different lysates with Mouse anti-EEA1 antibody (M1503-1) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Daudi cell lysate
    Lane 3: A431 cell lysate
    Lane 4: SH-SY5Y cell lysate
    Lane 5: NIH/3T3 cell lysate
    Lane 6: C6 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 162 kDa
    Observed band size: 162 kDa

    Exposure time: 10 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 (M1503-1) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human prostate tissue with Mouse anti-EEA1 antibody (<a href="/products/M1503-1" style="font-weight: bold;text-decoration: underline;">M1503-1</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/M1503-1" style="font-weight: bold;text-decoration: underline;">M1503-1</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human prostate tissue with Mouse anti-EEA1 antibody (M1503-1) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M1503-1) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse 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 ddH<sub>2</sub>O and PBS, and then probed with the antibody (<a href="/products/M1503-1" style="font-weight: bold;text-decoration: underline;">M1503-1</a>) at 1/400 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 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 (M1503-1) at 1/400 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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Citation