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重组RabMAb

重组Anti-Epsin 1抗体[EPR3023] (ab75879)

Reviews (1)Q&A (1)References (5)
Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
  • Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
  • Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
  • Immunocytochemistry/ Immunofluorescence - Anti-Epsin 1 antibody [EPR3023] (ab75879)
  • Immunocytochemistry/ Immunofluorescence - Anti-Epsin 1 antibody [EPR3023] (ab75879)
  • Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
  • Anti-Epsin 1 antibody [EPR3023] (ab75879)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3023] to Epsin 1
  • Suitable for: WB, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-Epsin 1抗体[EPR3023]
    参阅全部 Epsin 1 一抗

  • 描述

    兔单克隆抗体[EPR3023] to Epsin 1

  • 宿主

    Rabbit

  • 特异性

    The immunogen used for this product shares 100% homology with Epsin 2. Cross-reactivity with Epsin 2 has not been confirmed experimentally.

  • 经测试应用

    适用于: WB, ICC/IFmore details
    不适用于: IHC-P

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human Epsin 1 aa 450-550. The exact sequence is proprietary.
    Database link: Q9Y6I3

  • 阳性对照

    • ICC/IF: SH-SY5Y cells WB: SH-SY5Y and HeLa cell lysates

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab75879于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (1)
1/500 - 1/2000. Predicted molecular weight: 58 kDa.
ICC/IF
1/50.

For unpurified, use 1/100 - 1/250.
说明
WB
1/500 - 1/2000. Predicted molecular weight: 58 kDa.
ICC/IF
1/50.

For unpurified, use 1/100 - 1/250.
应用说明
Is unsuitable for IHC-P.

靶标

  • 功能

    Binds to membranes enriched in phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2). Modifies membrane curvature and facilitates the formation of clathrin-coated invaginations (By similarity). Regulates receptor-mediated endocytosis.

  • 序列相似性

    Belongs to the epsin family.
    Contains 1 ENTH (epsin N-terminal homology) domain.
    Contains 3 UIM (ubiquitin-interacting motif) repeats.

  • 结构域

    The NPF repeat domain is involved in EPS15 binding.
    The DPW repeat domain is involved in AP2A2 and clathrin binding.
    The [DE]-X(1,2)-F-X-X-[FL]-X-X-X-R motif mediates interaction with the AP-2 complex subunit AP2B1.

  • 翻译后修饰

    Phosphorylated on serine and/or threonine residues in mitotic cells. Phosphorylation reduces interaction with REPS2, AP-2 and the membrane fraction. Depolarization of synaptosomes results in dephosphorylation.
    Ubiquitinated.

  • 细胞定位

    Cytoplasm. Cell membrane. Nucleus. Membrane > clathrin-coated pit. Associated with the cytoplasmic membrane at sites where clathrin-coated pits are forming. Colocalizes with clathrin and AP-2 in a punctate pattern on the plasma membrane. Detected in presynaptic nerve terminals and in Golgi stacks. May shuttle to the nucleus when associated with ZBTB16/ZNF145.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • EH domain binding mitotic phosphoprotein antibody
    • EH domain-binding mitotic phosphoprotein antibody
    • EPN 1 antibody
    • Epn1 antibody
    • EPN1_HUMAN antibody
    • EPS 15 interacting protein 1 antibody
    • EPS-15-interacting protein 1 antibody
    • Epsin 1 antibody
    • Epsin-1 antibody
    see all

图片

  • Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Anti-Epsin 1 antibody [EPR3023] (ab75879) at 1/5000 dilution (purified) + rat brain at 20 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size: 58 kDa
    Observed band size: 94 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    All lanes : Anti-Epsin 1 antibody [EPR3023] (ab75879) at 1/1000 dilution (purified)

    Lane 1 : 293T cell lysate
    Lane 2 : K562 cell lysate
    Lane 3 : U-87 MG cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size: 58 kDa
    Observed band size: 94 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Anti-Epsin 1 antibody [EPR3023] (ab75879) at 1/5000 dilution (purified) + mouse brain tissue lysate at 20 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size: 58 kDa
    Observed band size: 94 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunocytochemistry/ Immunofluorescence - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Immunocytochemistry/ Immunofluorescence - Anti-Epsin 1 antibody [EPR3023] (ab75879)

    Immunofluorescence staining of HeLa cells with purified ab75879 at a working dilution of 1/50, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab75879 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunocytochemistry/ Immunofluorescence - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Immunocytochemistry/ Immunofluorescence - Anti-Epsin 1 antibody [EPR3023] (ab75879)

    Unpurified ab75879, at a 1/100 dilution, staining Epsin 1 in SH-SY5Y cells by Immunofluorescence.

  • Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Western blot - Anti-Epsin 1 antibody [EPR3023] (ab75879)
    All lanes : Anti-Epsin 1 antibody [EPR3023] (ab75879) at 1/2000 dilution (unpurified)

    Lane 1 : SH-SY5Y cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit-HRP at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 58 kDa

  • Anti-Epsin 1 antibody [EPR3023] (ab75879)
    Anti-Epsin 1 antibody [EPR3023] (ab75879)

文献 (5)

发表研究结果有使用 ab75879？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab75879 被引用在 5 文献中.

  • Kvalvaag A  et al. Clathrin mediates both internalization and vesicular release of triggered T cell receptor at the immunological synapse. Proc Natl Acad Sci U S A 120:e2211368120 (2023). PubMed: 36730202
  • Cabral-Dias R  et al. Fyn and TOM1L1 are recruited to clathrin-coated pits and regulate Akt signaling. J Cell Biol 221:N/A (2022). PubMed: 35238864
  • Wu N  et al. Proteomic characteristics of plasma and blood cells in natural aging rhesus monkeys. Proteomics 22:e2200049 (2022). PubMed: 36037246
  • Joseph JG  et al. Complimentary action of structured and unstructured domains of epsin supports clathrin-mediated endocytosis at high tension. Commun Biol 3:743 (2020). PubMed: 33293652
  • Costa R  et al. A transcriptional and post-transcriptional dysregulation of Dishevelled 1 and 2 underlies the Wnt signaling impairment in type I Gaucher disease experimental models. Hum Mol Genet 29:274-285 (2020). PubMed: 31816052

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-Epsin 1 antibody [EPR3023]

 Good
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (astrocytoma)
Loading amount
25 µg
Specification
astrocytoma
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

提交于 Apr 09 2009

Question

I write to ask you some information about your product N° ab75879 Anti-Epsin 1 antibody [EPR3023]. You indicate a predicted band size at 58KDa, but the Western blot image shows a band between 75 and 100KDa.   Coul you please explain me the reason of the difference between the predicted molecular weight and the real one?   Thank you in advance.   Best regards      

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Answer

Thank you for contacting us. I have quickly checked and found that this protein indeed runs at 80-90 kDa. The 58 kDa is the molecular weight of unprocessed raw protein which is calculated by combining the weight of amino acids as advised in Swissprot. 58kDa does not include the molecular weight of post translational changes of protein.   http://www.uniprot.org/uniprot/Q9Y6I3 I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. 

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