Validated using a knockout cell line
Recombinant
RabMAb

Anti-BIN1 antibody [EPR13463-25] (ab185950)

Overview

  • Product name
    Anti-BIN1 antibody [EPR13463-25]
    See all BIN1 primary antibodies
  • Description
    Rabbit monoclonal [EPR13463-25] to BIN1
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human BIN1 aa 400 to the C-terminus (C terminal).
    Database link: O00499

  • Positive control
    • U87-MG, Human fetal kidney and Hela cell lysates; Human fetal brain lysate;, Human skeletal muscle and mouse brain tissues; Hela and U87-MG cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab185950 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
Flow Cyt 1/90. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/100.
WB 1/1000. Detects a band of approximately 56, 65, 70, 90 kDa (predicted molecular weight: 65 kDa).
IHC-P 1/100.

Target

  • Function
    May be involved in regulation of synaptic vesicle endocytosis. May act as a tumor suppressor and inhibits malignant cell transformation.
  • Tissue specificity
    Ubiquitous. Highest expression in the brain and muscle. Isoform IIA is expressed only in the brain where it is concentrated in axon initial segments and nodes of Ranvier. Isoform BIN1 is widely expressed with highest expression in skeletal muscle.
  • Involvement in disease
    Defects in BIN1 are the cause of centronuclear myopathy autosomal recessive (ARCNM) [MIM:255200]; also known as autosomal recessive myotubular myopathy. Centronuclear myopathies are congenital muscle disorders characterized by progressive muscular weakness and wasting involving mainly limb girdle, trunk, and neck muscles. It may also affect distal muscles. Weakness may be present during childhood or adolescence or may not become evident until the third decade of life. Ptosis is a frequent clinical feature. The most prominent histopathologic features include high frequency of centrally located nuclei in muscle fibers not secondary to regeneration, radial arrangement of sarcoplasmic strands around the central nuclei, and predominance and hypotrophy of type 1 fibers.
  • Sequence similarities
    Contains 1 BAR domain.
    Contains 1 SH3 domain.
  • Post-translational
    modifications
    Phosphorylated by protein kinase C.
  • Cellular localization
    Cytoplasm and Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • AMPH 2 antibody
    • AMPH2 antibody
    • Amphiphysin 2 antibody
    • Amphiphysin II antibody
    • Amphiphysin like protein antibody
    • amphiphysin-like antibody
    • Amphiphysin-like protein antibody
    • AMPHL antibody
    • Bin1 antibody
    • BIN1_HUMAN antibody
    • Box Dependant MYC Interacting Protein 1 antibody
    • Box-dependent myc-interacting protein 1 antibody
    • Bridging integrator 1 antibody
    • DKFZp547F068 antibody
    • MGC10367 antibody
    • MGC105358 antibody
    • Myc box dependent interacting protein 1 antibody
    • Myc box-dependent-interacting protein 1 antibody
    • SH3P9 antibody
    see all

Images



  • Predicted band size : 65 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: BIN1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: U-87MG whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab185950 observed at 45-65 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab185950 was shown to specifically react with BIN1 when BIN1 knockout samples were used. Wild-type and BIN1 knockout samples were subjected to SDS-PAGE. Ab185950 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/10000 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.

  • Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling BIN1 with ab185950 at 1/100 dilution. Secondary ab: Ready to use HRP Polymer for Rabbit IgG. Counter stain: Hematoxylin.

  • Immunofluorescent analysis of HeLa cells labeling BIN1 with ab185950 at 1/100 dilution. Secondary ab: Goat anti rabbit IgG (Alexa Fluor®555) at 1/200 dilution. Fixative: -20℃ Acetone. Counter stain: Dapi (blue).

  • Flow cytometric analysis of Hela cells labeling BIN1 using ab185950 at 1/90 dilution (red). Secondary ab: Goat anti rabbit IgG (FITC) at 1/150 dilution. Fixative: 2% paraformaldehyde. Isotype control: Rabbit monoclonal IgG (green).

  • Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labeling BIN1 with ab185950 at 1/100 dilution. Secondary ab: Ready to use HRP Polymer for Rabbit IgG. Counter stain: Hematoxylin.

  • All lanes : Anti-BIN1 antibody [EPR13463-25] (ab185950) at 1/1000 dilution

    Lane 1 : U87-MG cell lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : Hela cell lysate
    Lane 4 : Human fetal brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 65 kDa

    Blocking buffer: 5% NFDM/TBST

  • Immunofluorescent analysis of U87-MG cells labeling BIN1 with ab185950 at 1/100 dilution. Secondary ab: Goat anti rabbit IgG (Alexa Fluor®555) at 1/200 dilution. Fixative: 4% paraformaldehyde. Counter stain: Dapi (blue)

References

ab185950 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
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Submitted Jun 07 2016

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