Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4245] to EEA1 - Early Endosome Marker
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-EEA1 antibody [EPR4245] - Early Endosome Marker
See all EEA1 primary antibodies
DescriptionRabbit monoclonal [EPR4245] to EEA1 - Early Endosome Marker
Tested applicationsSuitable for: ICC/IF, WBmore details
Unsuitable for: IP
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human EEA1 (N terminal). The exact sequence is proprietary.
Database link: Q15075
- WB: COS-1, NIH 3T3, C6, HeLa, Jurkat, Daudi, SH-SY5Y and JAR cell lysates. ICC/IF: JAR cells.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
- Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (Alexa Fluor® 488) (ab185039)
- Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (Alexa Fluor® 647) (ab196186)
- Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (Alexa Fluor® 594) (ab206913)
- Anti-EEA1 antibody [EPR4245] - BSA and Azide free (ab239942)
KO cell lines
KO cell lysates
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.
|ICC/IF||1/500 - 1/1000.|
|WB||1/10000 - 1/50000. Detects a band of approximately 170 kDa (predicted molecular weight: 162 kDa).|
FunctionBinds phospholipid vesicles containing phosphatidylinositol 3-phosphate and participates in endosomal trafficking.
Sequence similaritiesContains 1 C2H2-type zinc finger.
Contains 1 FYVE-type zinc finger.
DomainThe 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 localizationCytoplasm. Early endosome membrane.
- Information by UniProt
- Early endosome antigen 1 antibody
- Early endosome antigen 1, 162kD antibody
- Early endosome associated protein antibody
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 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : EEA1 knockout HeLa cell lysate
Lane 3 : Daudi cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 162 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?
Lanes 1- 4: Merged signal (red and green). Green - ab109110 observed at 175 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109110 was shown to react with EEA1 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab261822 (knockout cell lysate ab256897) lane below 175kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and EEA1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109110 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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.
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?
ab109110 has been referenced in 5 publications.
- Boivin M et al. Reduced autophagy upon C9ORF72 loss synergizes with dipeptide repeat protein toxicity in G4C2 repeat expansion disorders. EMBO J 39:e100574 (2020). PubMed: 31930538
- Yokoi A et al. Mechanisms of nuclear content loading to exosomes. Sci Adv 5:eaax8849 (2019). PubMed: 31799396
- Gillingham AK et al. In vivo identification of GTPase interactors by mitochondrial relocalization and proximity biotinylation. Elife 8:N/A (2019). PubMed: 31294692
- Wolff NA et al. Evidence for mitochondrial localization of divalent metal transporter 1 (DMT1). FASEB J 28:2134-45 (2014). PubMed: 24448823
- Choi S et al. Globotriaosylceramide induces lysosomal degradation of endothelial KCa3.1 in fabry disease. Arterioscler Thromb Vasc Biol 34:81-9 (2014). PubMed: 24158513