Anti-Epsin 1 antibody [EPR3023] (ab75879)

Overview

  • Product nameAnti-Epsin 1 antibody [EPR3023]
    See all Epsin 1 primary antibodies
  • Description
    Rabbit monoclonal [EPR3023] to Epsin 1
  • Tested applicationsSuitable for: WB, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Epsin 1 aa 450-550.
    Database link: Q9Y6I3

  • Positive control
    • ICC/IF: SH-SY5Y cells WB: SH-SY5Y and HeLa cell lysates
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Properties

Applications

Our Abpromise guarantee covers the use of ab75879 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/2000. Predicted molecular weight: 58 kDa.
ICC/IF 1/50.

For unpurified, use 1/100 - 1/250.

  • Application notesIs unsuitable for IHC-P.
  • Target

    • FunctionBinds 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.
    • Sequence similaritiesBelongs to the epsin family.
      Contains 1 ENTH (epsin N-terminal homology) domain.
      Contains 3 UIM (ubiquitin-interacting motif) repeats.
    • DomainThe 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.
    • Post-translational
      modifications
      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.
    • Cellular localizationCytoplasm. 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
    • Database links
    • Alternative names
      • 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

    Anti-Epsin 1 antibody [EPR3023] images

    • 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

    • 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 : U87-MG cell lysate

      Lysates/proteins at 10 µg per lane.

      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

    • Anti-Epsin 1 antibody [EPR3023] (ab75879) at 1/5000 dilution (purified) + mouse brain 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

    • 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.

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

    • 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
      Goat anti-rabbit-HRP at 1/1000 dilution
      Developed using the ECL technique

      Predicted band size : 58 kDa

    References for Anti-Epsin 1 antibody [EPR3023] (ab75879)

    ab75879 has not yet been referenced specifically in any publications.

    Product Wall

    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 Swisspr...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    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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    Verified customer

    Submitted Apr 09 2009

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"