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Immunocytochemistry - EEA1 antibody - Early Endosome Marker (ab2900)
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Immunocytochemistry/ Immunofluorescence - EEA1 antibody - Early Endosome Marker (ab2900)
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Immunocytochemistry - EEA1 antibody - Early Endosome Marker (ab2900)
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Product Name
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EEA1 antibody - Early Endosome Marker
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See all EEA1 - Early Endosome Marker antibodies (6)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to EEA1 - Early Endosome Marker
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Immunogen
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Synthetic peptide derived from within residues 1350 to the C-terminus of Human EEA1.
(Peptide available as ab14946.)
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Reacts with
(species key)
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Hu, Ms, Rat, Dog, Hm, Xl
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Specificity
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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.
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Tested applications
(see key)
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ICC/IF, IHC (Methanol fixed), IHC-FoFr, IHC-P, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC: 1/200 - 1/500.
ICC/IF: Use at a concentration of 1 µg/ml.
Immunohistochemistry (Methanol fixed cells): Use at an assay dependent dilution.
IHC-P: 1/500.
IHC-FoFr: Use at an assay dependent dilution (PMID 18852301).
IP: Use at an assay dependent dilution.
WB: Use at 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).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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Rat brain
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Cellular localization
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Endosome
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Research areas
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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
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Relevance
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Early endosomal antigen 1 (EEA1) is a 162 kDa membrane bound protein component specific to the early endosome and is essential for fusion between early endocytic vesicles. Early endosomes are cytoplasmic compartments which fuse with endocytic vesicles for redistribution of extracellular compounds to alternate destinations. Zinc finger like domains, similar to those found in nucleic acid binding proteins, are located in the amino and carboxyl terminal domains of EEA1. The carboxyl terminal zinc finger like domain is conserved in several other non nuclear proteins, some of which are also involved in intracellular protein trafficking. In addition, this domain is an authentic zinc binding domain and is critical to the intracellular localization of EEA1. Membrane association of EEA1 has been shown to be dependent on phosphatidyl 3 kinase activity; inhibitors of this enzyme cause EEA1 to dissociate from early endosomes.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.10 - 0.80 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this EEA1 antibody - Early Endosome Marker is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for EEA1 antibody - Early Endosome Marker images, references, products related to ab2900 and other tools.
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EEA1 antibody - Early Endosome Marker images:
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 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
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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)
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
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Immunofluorescence - EEA1 antibody - Early Endosome Marker (ab2900)
Immunolocalisation of EEA1 in HeLa cells using ab2900.
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Western blot - EEA1 antibody - Early Endosome Marker (ab2900)
All lanes : 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. We are unsure as to the identity of these extra bands.
Fluorescence detection of secondary antibody.
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- EEA1 antibody - Early Endosome Marker (ab2900)
All lanes : 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 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?)
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Western blot - EEA1 antibody - Early Endosome Marker (ab2900)
All lanes : 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 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.
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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
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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
See Abreview
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References for EEA1 antibody - Early Endosome Marker (ab2900)
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This product has been used in: (two most recent references)
Ha SD et al. Cathepsin B-mediated autophagy flux facilitates the anthrax toxin receptor 2-mediated delivery of anthrax lethal factor into the cytoplasm. J Biol Chem 285:2120-9 (2010). WB; Human. PubMed: 19858192
Tsitoura E et al. Infection with herpes simplex type 1-based amplicon vectors results in an IRF3/7-dependent, TLR-independent activation of the innate antiviral response in primary human fibroblasts. J Gen Virol 90:2209-20 (2009). PubMed: 19515829
See all 25 publication references for this product.
If you publish research using ab2900 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to EEA1
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