Anti-EEA1抗体- Early Endosome Marker (ab2900)
- 数据表
- 具体的参考文献 (77)
- 实验方案
概述
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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
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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.
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经测试应用适用于: ICC/IF, IHC (Methanol fixed), IHC-P, WB, ICC, IP, IHC-FoFrmore details
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种属反应性与反应: Mouse, Rat, Chicken, Hamster, Cow, Dog, Human, Xenopus laevis, Zebrafish, Rhesus monkey, Aplysia
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免疫原
Synthetic peptide derived from within residues 1350 to the C-terminus of Human EEA1.
(Peptide available as ab14946.)
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阳性对照
- Rat brain
性能
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形式Liquid
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存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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存储溶液Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
纯度Immunogen affinity purified
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克隆多克隆
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同种型IgG
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研究领域
相关产品
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Immunohistochemistry kits
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Positive Controls
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Related Products
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). Abcam recommends using milk as the blocking agent. |
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| ICC | 1/200 - 1/500. | |
| IP | Use at an assay dependent concentration. | |
| IHC-FoFr | Use at an assay dependent concentration. |
靶标
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功能Binds phospholipid vesicles containing phosphatidylinositol 3-phosphate and participates in endosomal trafficking.
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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.
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细胞定位Cytoplasm. Early endosome membrane.
- Information by UniProt
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数据库链接
- 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 -
别名
- Early endosome antigen 1 antibody
- Early endosome antigen 1, 162kD antibody
- Early endosome associated protein antibody
see all
Anti-EEA1 antibody - Early Endosome Marker 图像
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Immunocytochemistry/ Immunofluorescence - Anti-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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Western blot - Anti-EEA1 antibody - Early Endosome Marker (ab2900)All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human) Whole Cell Lysate
Lane 3 : NIH 3T3 (Mouse) Whole Cell Lysate
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : HEK293 (Human) Whole Cell Lysate
Lane 6 : NIH 3T3 (Mouse) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG H&L (HRP) at 1/50000 dilution
Developed using the ECL technique
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.
Exposure time : 30 secondsThis blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin (Lanes 1-3) or 3% Milk (Lanes 4-6) before being incubated with ab2900 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
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Western blot - Anti-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. -
Immunohistochemistry (Paraffin-embedded sections) - Anti-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 - Anti-EEA1 antibody - Early Endosome Marker (ab2900)This image is courtesy of 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-β.
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Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody - Early Endosome Marker (ab2900)This image is courtesy of 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.
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Immunocytochemistry/ Immunofluorescence - Anti-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.
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Western blot - Anti-EEA1 antibody - Early Endosome Marker (ab2900)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 withHuman EEA1 peptide (ab14946) at 1 µg
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 : 180 kDa (why is the actual band size different from the predicted?)
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?) -
Western blot - Anti-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 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 secondsThe 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.
实验方案
Anti-EEA1 antibody - Early Endosome Marker (ab2900)参考文献
This product has been referenced in:
- Kleinecke S et al. Peroxisomal dysfunctions cause lysosomal storage and axonal Kv1 channel redistribution in peripheral neuropathy. Elife 6:N/A (2017). IF ; Mouse . Read more (PubMed: 28470148) »
- Ortiz DA et al. Protein-Protein Interactions Suggest Novel Activities of Human Cytomegalovirus Tegument Protein pUL103. J Virol 90:7798-810 (2016). WB, ICC/IF ; Human . Read more (PubMed: 27334581) »
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