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Products:Neuroscience >> Cell Type Marker >> Neuron marker >> Soma marker
Overview
Product name
Anti-EEA1 antibody - Early Endosome Marker
See all EEA1 products (9) ...
Description
Rabbit polyclonal to EEA1 - Early Endosome Marker
Specificity
Detects 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 applications
ICC/IF, IHC (Methanol fixed), IHC-P, WB, ICC, IP, IHC-FoFrmore details
Cross reactivity
Reacts with
Mouse, Rat, Hamster, Cow, Dog, Human, Xenopus laevis
Immunogen
Synthetic peptide derived from within residues 1350 to the C-terminus of Human EEA1.
(Peptide available as ab14946.)
Positive control
Rat brain
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Neuroscience >> Neurotransmission >> Intracellular Signaling >> Regulation
Signal Transduction >> Protein Trafficking >> Organelle Proteins
Tags & Cell Markers >> Subcellular Markers >> Organelles >> Endosome
Neuroscience >> Cell Type Marker >> Neuron marker >> Soma marker
Applications
Show applications key
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.
ICC/IF: Use a concentration of 1 µg/ml
IHC (Methanol fixed): Use at an assay dependent dilution.
IHC-P: 1/500
WB: Use a concentration of 1 µg/mlDetects a band of approximately 180 kDa (predicted molecular weight: 160 kDa).Can be blocked with EEA1 peptide (ab14946).
ICC: 1/200 - 1/500.
IP: Use at an assay dependent dilution.
IHC-FoFr: Use at an assay dependent dilution.
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.
Target information above from: UniProt accessionQ15075
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Early endosome antigen 1 antibody
- Early endosome antigen 1, 162kD antibody
- Early endosome associated protein antibody
see all
Database links
- Entrez Gene: 8411 Human
- Entrez Gene: 216238 Mouse
- Entrez Gene: 314764 Rat
- Omim: 605070 Human
- SwissProt: Q15075 Human
- SwissProt: Q8BL66 Mouse
- Unigene: 567367 Human
- Unigene: 210035 Mouse
see all
Anti-EEA1 antibody - Early Endosome Marker images:
Immunocytochemistry - EEA1 antibody - Early Endosome Marker (ab2900)
Confocal 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.
Randal Moldrich, CNRS UMR7637, ESPCI, France
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).
Immunocytochemistry - EEA1 antibody - Early Endosome Marker (ab2900)
ab2900 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.
Randal Moldrich, CNRS UMR7637, ESPCI, France
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.
Fluorescence detection of secondary antibody.
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 with EEA1 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
The Western blot shows that ab14944 reacts strongly with mouse 3T3 and CHO cell lysates. Weak cross-reactivity is seen with Xenopus, mouse brain, mouse liver, rat brain and dog lysates. The antibody does not appear to cross-react with rat liver lysate.
Immunohistochemistry (Paraffin-embedded sections) - EEA1 antibody - Early Endosome Marker (ab2900)
Image courtesy of Human Protein Atlas
Paraffin 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
Immunocytochemistry/ Immunofluorescence - EEA1 antibody - Early Endosome Marker (ab2900)
ab2900 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.
This image is courtesy of an anonymous Abreview
References for Anti-EEA1 antibody - Early Endosome Marker (ab2900)
This product has been referenced in:
- Grimm MOet al. Docosahexaenoic Acid Reduces Amyloid {beta} Production via Multiple Pleiotropic Mechanisms. J Biol Chem 286:14028-39 (2011). WB; Human.Read more (PubMed: 21324907) »
- Giudice Jet al. Differential endocytosis and signaling dynamics of insulin receptor variants IR-A and IR-B. J Cell Sci 124:801-11 (2011).Read more (PubMed: 21303927) »
See all 33 publications for this product
Publishing research using ab2900? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"