Anti-EEA1抗体- Early Endosome Marker (ab2900)

概述

  • 产品名称Anti-EEA1抗体- Early Endosome Marker
    参阅全部 EEA1 一抗
  • 描述
    兔多克隆抗体to EEA1 - Early Endosome Marker
  • 特异性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.
  • 经测试应用适用于: ICC/IF, IHC (Methanol fixed), IHC-P, WB, ICC, IP, IHC-FoFrmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Hamster, Cow, Dog, Human, Xenopus laevis, Zebrafish, Rhesus monkey, Aplysia
  • 免疫原

    Synthetic peptide derived from within residues 1350 to the C-terminus of Human EEA1.

    (Peptide available as ab14946.)

  • 阳性对照
    • Rat brain

性能

应用

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.

应用 Ab评论 说明
ICC/IF Use a concentration of 1 µg/ml.
IHC (Methanol fixed) Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 160 kDa).Can be blocked with Human EEA1 peptide (ab14946).
ICC 1/200 - 1/500.
IP Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration.

靶标

  • 功能Binds phospholipid vesicles containing phosphatidylinositol 3-phosphate and participates in endosomal trafficking.
  • 序列相似性Contains 1 C2H2-type zinc finger.
    Contains 1 FYVE-type zinc finger.
  • 结构域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.
  • 细胞定位Cytoplasm. Early endosome membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Early endosome antigen 1 antibody
    • Early endosome antigen 1, 162kD antibody
    • Early endosome associated protein antibody
    • EEA 1 antibody
    • EEA1 antibody
    • EEA1_HUMAN antibody
    • Endosome associated protein p162 antibody
    • Endosome-associated protein p162 antibody
    • MST105 antibody
    • MSTP105 antibody
    • ZFYVE2 antibody
    • Zinc finger FYVE domain containing protein 2 antibody
    • Zinc finger FYVE domain-containing protein 2 antibody
    see all

Anti-EEA1 antibody - Early Endosome Marker 图像

  • 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).

  • 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.
  • 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

  • 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-β.

  • 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.

  • 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.

    See Abreview



  • Performed under reducing conditions.

    Predicted band size : 160 kDa
    Lane 1 - 2 : EEA1 antibody - Early Endosome Marker (ab2900) at 1/500 dilution, HEK293 Whole Cell lysate at 20 ug Lane 1 : as above Lane 2 : EEA1 peptide (ab14946) at 1 ug Secondary Goat polyclonal to Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution Performed under reducing conditions. Predicted band size : 160kD Observed band size : 180kD (why is the actual band size different from the predicted?)
  • 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 Anti-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.

Anti-EEA1 antibody - Early Endosome Marker (ab2900)参考文献

This product has been referenced in:
  • Bourseau-Guilmain E  et al. Hypoxia regulates global membrane protein endocytosis through caveolin-1 in cancer cells. Nat Commun 7:11371 (2016). WB, IHC-Fr, ICC/IF . Read more (PubMed: 27094744) »
  • Raha AA  et al. Neuroprotective Effect of TREM-2 in Aging and Alzheimer's Disease Model. J Alzheimers Dis N/A:N/A (2016). IHC ; Mouse . Read more (PubMed: 27662313) »

See all 65 Publications for this product

Product Wall

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Temperature: 20°C
Antigen retrieval step None
Sample Aplysia Tissue sections (buccal ganglia)
Specification buccal ganglia
Permeabilization Yes - triton 1.5%
Fixative Paraformaldehyde
Username

Ms. Francoise Geffroy

Verified customer

提交于 Dec 06 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Caco-2 cell)
Specification Caco-2 cell
Fixative Paraformaldehyde
Permeabilization Yes - 0.25% Triton X-100 in TBS
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Mar 27 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Acinar epithelial explants in collagen)
Permeabilization Yes - TritonX100 0.025% 10 min RT
Specification Acinar epithelial explants in collagen
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 19°C
Fixative Paraformaldehyde
Username

Dr. Clara Lubeseder-Martellato

Verified customer

提交于 Jun 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Pancreas or PDAC cells)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 30 µg
Specification Pancreas or PDAC cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 19°C
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Verified customer

提交于 Jun 14 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (neuron)
Permeabilization Yes - 0.2%Triton
Specification neuron
Blocking step Serum as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative Formaldehyde
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Abcam user community

Verified customer

提交于 Nov 30 2015

Application Western blot
Sample Mouse Tissue lysate - whole (BRAIN)
Gel Running Conditions Reduced Denaturing (8%)
Loading amount 50 µg
Treatment alpha synuclein
Specification BRAIN
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Abcam user community

Verified customer

提交于 Nov 09 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Sample Chicken Cell (Primary oculomotor culture)
Specification Primary oculomotor culture
Permeabilization Yes - PBS + 0.1% Tween20
Fixative Formaldehyde
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Abcam user community

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提交于 Oct 27 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Human Cell (ovarian carcinoma TOV112-D cell line)
Specification ovarian carcinoma TOV112-D cell line
Permeabilization Yes - 0.1% triton X for 5 min at RT
Fixative Paraformaldehyde
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提交于 Aug 27 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 22°C
Sample Rat Cell (hippocampal neuron)
Specification hippocampal neuron
Permeabilization Yes - 2% TritonX100
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 13 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Goat serum 5% + BSA 1% as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample Human Cell (U87 Glioblastoma)
Specification U87 Glioblastoma
Permeabilization Yes - triton 0.2%
Fixative Paraformaldehyde
Username

Ms. Francoise Geffroy

Verified customer

提交于 Aug 01 2014

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