Anti-EEA1 antibody - Early Endosome Marker (ab2900)
Overview
- Product nameAnti-EEA1 antibody - Early Endosome MarkerSee all EEA1 primary antibodies ...
- DescriptionRabbit polyclonal to EEA1 - Early Endosome Marker
- SpecificityDetects a band at 180kDa that represents EEA1 in Western blotting on human cell lines (corresponds to results seen in Mu et al). Also detects a band at 100kDa, we are unsure as to the identity of this band. Immunofluorescence staining of EEA1 in HeLa cells yields a punctate staining pattern consistent with the cytoplasmic distribution of endosomes.
- Tested applicationsICC/IF, IHC (Methanol fixed), IHC-P, WB, ICC, IP, IHC-FoFr more details
- Species reactivityReacts with: Mouse, Rat, Hamster, Cow, Dog, Human, Xenopus laevis, Zebrafish, Rhesus monkey
- Immunogen
Synthetic peptide derived from within residues 1350 to the C-terminus of Human EEA1.
(Peptide available as ab14946.)
- Positive controlRat brain
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab2900 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| ICC/IF | ICC/IF: Use a concentration of 1 µg/ml. |
| IHC (Methanol fixed) | IHC (Methanol fixed): Use at an assay dependent dilution. |
| IHC-P | IHC-P: Use at an assay dependent dilution. |
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 160 kDa).Can be blocked with EEA1 peptide (ab14946). |
| ICC | ICC: 1/200 - 1/500. |
| IP | IP: Use at an assay dependent dilution. |
| IHC-FoFr | IHC-FoFr: Use at an assay dependent dilution. |
Target
- 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.
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Database links
- Entrez Gene: 8411 Human
- Entrez Gene: 216238 Mouse
- Entrez Gene: 314764 Rat
- Omim: 605070 Human
- SwissProt: Q15075 Human
- SwissProt: H9G0K4 Human
- SwissProt: Q8BL66 Mouse
- SwissProt: H9G0K4 Rhesus monkey
- Unigene: 567367 Human
- Unigene: 210035 Mouse
- Unigene: 23513 Rat
see all
Target information above from: UniProt accession
Q15075
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- Early endosome antigen 1 antibodyEarly endosome antigen 1, 162kD antibodyEarly endosome associated protein antibody
- EEA 1 antibodyEEA1 antibodyEEA1_HUMAN antibodyEndosome associated protein p162 antibodyEndosome-associated protein p162 antibodyMST105 antibodyMSTP105 antibodyZFYVE2 antibodyZinc finger FYVE domain containing protein 2 antibodyZinc finger FYVE domain-containing protein 2 antibody
see all
Anti-EEA1 antibody - Early Endosome Marker images
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Immunocytochemistry - EEA1 antibody - Early Endosome Marker (ab2900)Randal Moldrich, CNRS UMR7637, ESPCI, FranceConfocal microscopy of fixed primary cultures of rat hippocampal neurons (embryonic day 18) showing immunocytochemical labelling of rabbit polyclonal to EEA1 (ab2900, 1/200; Alexa Fluor 488 1/200; green) and monoclonal mouse anti-b-tubulin-II (1/400; Alexa Fluor 568 1/200; red). Magnification 63x ; insert 180x. -
Immunocytochemistry/ Immunofluorescence - EEA1 antibody - Early Endosome Marker (ab2900)ICC/IF image of ab2900 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab2900, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Immunocytochemistry - EEA1 antibody - Early Endosome Marker (ab2900)Randal Moldrich, CNRS UMR7637, ESPCI, Franceab2900 Rabbit polyclonal to EEA1 (1/500) was used in fixed HEK cells that had been transfected with Rab5-GFP. Fluorescence microscopy of fixed Rab5-transfected HEK cells showing (left to right) immunocytochemical labelling rabbit polyclonal to EEA1 ab2900 (1/500; Alexa Fluor 568; red), and Rab5-GFP (green) with their overlay (far right). Scale bar = 5µm. Showing little co-localisation as expected. Immunofluorescence microscopy revealed a typically punctuate staining of EEA1 in the HEK cells.
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Immunofluorescence - EEA1 antibody - Early Endosome Marker (ab2900)Immunolocalisation of EEA1 in HeLa cells using ab2900. -
Western blot - EEA1 antibody - Early Endosome Marker (ab2900)All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1/1000 dilution
Lane 1 : HeLa nuclear
Lane 2 : HeLa whole cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : HEK293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Alexa Fluor anti rabbit at 1/50000 dilution
Performed under reducing conditions.
Predicted band size : 160 kDa
Observed band size : 180 kDa (why is the actual band size different from the predicted?)
Additional bands at : 100 kDa,41 kDa,50 kDa. We are unsure as to the identity of these extra bands. -
Western blot - EEA1 peptide (ab14946)All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1/500 dilution
Lane 1 : HEK293 Whole Cell lysate
Lane 2 : HEK293 Whole Cell lysate withEEA1 peptide (ab14946) at 1 µg
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size : 160 kDa
Observed band size : 180 kDa (why is the actual band size different from the predicted?) -
Western blot - EEA1 antibody - Early Endosome Marker (ab2900)All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1 µg/ml
Lane 1 : Xenopus laevis lysate
Lane 2 : Mouse 3T3 cell lysate
Lane 3 : Mouse brain cell lysate
Lane 4 : Mouse liver cell lysate
Lane 5 : Rat brain cell lysate
Lane 6 : Rat liver cell lysate
Lane 7 : Dog lysate
Lane 8 : CHO cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size : 160 kDa
Observed band size : 100,180 kDa (why is the actual band size different from the predicted?)
Exposure time : 30 seconds -
Immunohistochemistry (Paraffin-embedded sections) - EEA1 antibody - Early Endosome Marker (ab2900)Image courtesy of Human Protein AtlasParaffin embedded sections of human prostate tissue were incubated with ab2900 (1/500 dilution) at room temperature for 30 mins. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab2900 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further images can be found at www.proteinatlas.org
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Immunocytochemistry/ Immunofluorescence - EEA1 antibody - Early Endosome Marker (ab2900)This image is courtesy of an anonymous Abreviewab2900 staining mouse L-cells by ICC/IF. Cells were formaldehyde fixed, permeabilized in Triton X-100 and incubated with ab2900 diluted 1/1000 for 1 hour at 37°C. A Cy2® conjugated goat anti-rabbit antibody was used as the secondary.
References for Anti-EEA1 antibody - Early Endosome Marker (ab2900)
This product has been referenced in:
- Abramowski D et al. Transgenic expression of intraneuronal Aß42 but not Aß40 leads to cellular Aß lesions, degeneration, and functional impairment without typical Alzheimer's disease pathology. J Neurosci 32:1273-83 (2012). ICC/IF ; Mouse . Read more (PubMed: 22279212) »
- Xu Y et al. Epidermal Growth Factor-induced Vacuolar (H+)-ATPase Assembly: A ROLE IN SIGNALING VIA mTORC1 ACTIVATION. J Biol Chem 287:26409-22 (2012). ICC/IF . Read more (PubMed: 22689575) »
