Anti-EEA1抗体- Early Endosome Marker (ab2900)
Key features and details
- Rabbit polyclonal to EEA1 - Early Endosome Marker
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Dog, Human, Xenopus laevis, Chinese hamster
- Isotype: IgG
选择批间可重复性更高的重组抗体
- 研究可靠 —— 各批次间结果一致且可重复
- 长期批量供应 —— 采用重组技术,可实现快速生产
- 首次实验即可成功 —— 经过大量验证确认了特异性
- 符合伦理标准 —— 产品不含动物成分
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 -
宿主
Rabbit -
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, WBmore details -
种属反应性
与反应: Mouse, Rat, Dog, Human, Xenopus laevis, Chinese hamster
预测可用于: Chicken, Hamster, Cow, Zebrafish, Rhesus monkey, Aplysia -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- ICC: HepG2 cells
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常规说明
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
性能
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形式
Liquid -
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. -
存储溶液
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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纯度
Immunogen affinity purified -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
应用
应用 | Ab评论 | 说明 |
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ICC/IF | (23) |
Use a concentration of 1 - 5 µg/ml.
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WB | (12) |
Use a concentration of 1 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 160 kDa).
Abcam recommends using milk as the blocking agent. |
说明 |
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ICC/IF
Use a concentration of 1 - 5 µg/ml. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 160 kDa). Abcam recommends using milk as the blocking agent. |
靶标
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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
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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
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别名
- Early endosome antigen 1 antibody
- Early endosome antigen 1, 162kD antibody
- Early endosome associated protein antibody
see all
图片
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ab2900 staining EEA1 in HepG2 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab2900 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: EEA1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: NIH3T3 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab2900 observed at 162 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab2900 was shown to recognize EEA1 in wild-type HAP1 cells as signal was lost at the expected MW in EEA1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and EEA1 knockout samples were subjected to SDS-PAGE. Ab2900 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lane 4 : HeLa whole cell lysate
Lane 5 : HEK-293 whole cell lysate
Lane 6 : NIH 3T3 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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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All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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. -
Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody - Early Endosome Marker (ab2900)This image is courtesy of Randal Moldrich, CNRS UMR7637, ESPCI, France
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.
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Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody - Early Endosome Marker (ab2900)This image is courtesy of an anonymous Abreview
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.
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All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1/500 dilution
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HEK293 Whole Cell lysate with Human EEA1 peptide (ab14946) at 1 µg
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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?) -
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
All lanes : 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
数据表及文件
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SDS download
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Datasheet download
文献 (239)
ab2900 被引用在 239 文献中.
- Millarte V et al. Rabaptin5 targets autophagy to damaged endosomes and Salmonella vacuoles via FIP200 and ATG16L1. EMBO Rep 23:e53429 (2022). PubMed: 34704340
- Shrivastava R et al. Rabaptin5 acts as a key regulator for Rab7l1-mediated phagosome maturation process. Immunology 165:328-340 (2022). PubMed: 34888849
- Barthet VJA et al. DRAM-4 and DRAM-5 are compensatory regulators of autophagy and cell survival in nutrient-deprived conditions. FEBS J 289:3752-3769 (2022). PubMed: 35060334
- Wang F et al. The p97 Inhibitor UPCDC-30245 Blocks Endo-Lysosomal Degradation. Pharmaceuticals (Basel) 15:N/A (2022). PubMed: 35215314
- Governa V et al. Landscape of surfaceome and endocytome in human glioma is divergent and depends on cellular spatial organization. Proc Natl Acad Sci U S A 119:N/A (2022). PubMed: 35217608