Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)

Overview

  • Product name

    Anti-EEA1 antibody [EPR4245] - Early Endosome Marker
    See all EEA1 primary antibodies
  • Description

    Rabbit monoclonal [EPR4245] to EEA1 - Early Endosome Marker
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human EEA1 (N terminal). The exact sequence is proprietary.
    Database link: Q15075

  • Positive control

    • WB: COS-1, NIH 3T3, C6, HeLa, Jurkat and JAR cell lysates. ICC/IF: JAR cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109110 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/500 - 1/1000.
WB 1/10000 - 1/50000. Detects a band of approximately 170 kDa (predicted molecular weight: 162 kDa).
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Binds phospholipid vesicles containing phosphatidylinositol 3-phosphate and participates in endosomal trafficking.
    • Sequence similarities

      Contains 1 C2H2-type zinc finger.
      Contains 1 FYVE-type zinc finger.
    • Domain

      The FYVE-type zinc finger domain mediates interactions with phosphatidylinositol 3-phosphate in membranes of early endosomes and penetrates bilayers. The FYVE domain insertion into PtdIns(3)P-enriched membranes is substantially increased in acidic conditions.
    • Cellular localization

      Cytoplasm. Early endosome membrane.
    • Information by UniProt
    • Database links

    • Alternative names

      • 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
      see all

    Images

    • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: Early Endosome Marker knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: NIH3T3 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab109110 observed at 162 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab109110 was shown to recognize Early Endosome Marker in wild-type HAP1 cells as signal was lost at the expected MW in Early Endosome Marker knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Early Endosome Marker knockout samples were subjected to SDS-PAGE. Ab109110 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/Immunofluorescence analysis of JAR (human placenta choriocarcinoma epithelial) cells labelling EEA1 with ab109110 at a dilution of 1/250. Cells were fixed with 4% paraformaldehye and permeabilized with 0.1% TritonX-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG was used as the secondary antibody at a dilution of 1/1000. Counterstained with DAPI and ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), at a dilution of 1/200.

      Image shows cytoplasmic staining in JAR cell line.

    • All lanes : Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110) at 1/10000 dilution

      Lane 1 : COS-1 cell lysate
      Lane 2 : NIH 3T3 cell lysate
      Lane 3 : C6 cell lysate
      Lane 4 : HeLa cell lysate
      Lane 5 : Jurkat cell lysate
      Lane 6 : JAR cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 162 kDa
      Observed band size: 170 kDa
      why is the actual band size different from the predicted?

    References

    This product has been referenced in:

    • Gillingham AK  et al. In vivo identification of GTPase interactors by mitochondrial relocalization and proximity biotinylation. Elife 8:N/A (2019). Read more (PubMed: 31294692) »
    • Wolff NA  et al. Evidence for mitochondrial localization of divalent metal transporter 1 (DMT1). FASEB J 28:2134-45 (2014). Read more (PubMed: 24448823) »
    See all 3 Publications for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Specification
    HEK cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

    Abcam user community

    Verified customer

    Submitted Jun 14 2019

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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