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 and JAR cell lysates. ICC/IF: JAR cells.
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, 0.05% BSA, 50% Tissue culture supernatant
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)
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 1/10000 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?
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) »