Anti-EEA1 antibody [1G11] - Early Endosome Marker (ab70521)

Overview

  • Product name
    Anti-EEA1 antibody [1G11] - Early Endosome Marker
    See all EEA1 primary antibodies
  • Description
    Mouse monoclonal [1G11] to EEA1 - Early Endosome Marker
  • Tested applications
    Suitable for: ICC/IFmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit
  • Immunogen

    Recombinant fragment derived from within residues 50 - 150 of Human EEA1.

  • Positive control
    • This anitbody gave a positive immunofluorescent signal in the following cell lines: HeLa HepG2
  • General notes

    This antibody clone is manufactured by Abcam.

Properties

Applications

Our Abpromise guarantee covers the use of ab70521 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 Use a concentration of 1 - 5 µg/ml.
  • Application notes
    Is unsuitable for WB.
  • 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

    • ICC/IF image of ab70521 stained HepG2 cells. The cells were fixed with 4% PFA (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70521 at 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa and MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, HepG2 and MCF7 cells at 5µg/ml.
    • ICC/IF image of ab70521 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70521, 10µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 oat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • ab70521 staining EEA1 in human THP-1 cells by Immunocytochemistry/ immunofluorescence. Cells were formaldehyde fixed, permeabilized in TritonX100 and blocked in 1% BSA for 1 hour at 25°C. The primary antibody was diluted 1/200 (PBS + Tween20 + BSA 1%) and incubated with sample for 1 hour at 25°C. A 1.4 nm Gold (ultra-small) and Alexa Fluor® 594 conjugated goat polyclonal to mouse IgG1, diluted 1/500 was used as secondary. The red staining in image show EEA1 staining whilst green cytosolic staining corresponds to StarD5.

      See Abreview

    References

    This product has been referenced in:
    • Poulsen ET  et al. An Aberrant Phosphorylation of Amyloid Precursor Protein Tyrosine Regulates Its Trafficking and the Binding to the Clathrin Endocytic Complex in Neural Stem Cells of Alzheimer's Disease Patients. Front Mol Neurosci 10:59 (2017). ICC/IF . Read more (PubMed: 28360834) »
    • Zollo A  et al. Sortilin-Related Receptor Expression in Human Neural Stem Cells Derived from Alzheimer's Disease Patients Carrying the APOE Epsilon 4 Allele. Neural Plast 2017:1892612 (2017). ICC/IF ; Human . Read more (PubMed: 28634550) »

    See all 40 Publications for this product

    Customer reviews and Q&As

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Pig Cell (Epithelial intestinal cell line)
    Permeabilization
    Yes - 0.1% Tween
    Specification
    Epithelial intestinal cell line
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Formaldehyde
    Username

    Sébastien Holbert

    Verified customer

    Submitted Aug 22 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: 24°C
    Sample
    Monkey Cell (Kidney)
    Specification
    Kidney
    Permeabilization
    Yes - TritonX100 al 0,1%, Tween20
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Oct 24 2014

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Sample
    Mouse Cell (hippocampal neuronal culture)
    Specification
    hippocampal neuronal culture
    Permeabilization
    Yes - 0.2% Triton
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Sep 12 2013

    Thank you for your inquiry.

    We have two secondary antibodies that match this mouse IgG1 primary antibody:

    ab97239:

    http://www.abcam.com/index.html?datasheet=97239 (or use the following: http://www.abcam.com/index.html?data...

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Guinea pig Cell (distal colon)
    Specification
    distal colon
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 19 2010

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Brain)
    Specification
    Brain
    Username

    Dr. Sophie Pezet

    Verified customer

    Submitted Sep 08 2010

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry
    Sample
    Mouse Cell (primary hippocampal neurons)
    Specification
    primary hippocampal neurons
    Fixative
    4% PFA/Sucrose
    Permeabilization
    Yes - 0,2% Triton X-100
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Jun 10 2010

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (THP-1)
    Specification
    THP-1
    Fixative
    Formaldehyde
    Permeabilization
    Yes - Triton X-100
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Apr 01 2010

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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