Validated using a knockout cell line
Recombinant
RabMAb

Anti-BIN1 antibody [EPR13463] (ab182562)

Overview

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

    Recombinant fragment within Human BIN1 aa 400 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: O00499

  • Positive control
    • A431, HeLa, Human fetal kidney, Human skeletal muscle, Human fetal brain lysates; Human skeletal muscle, Human clear cell carcinoma of kidney and Human brain tissues; U87-MG and HeLa cells; U87-MG cell lysate.
  • 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 ab182562 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 1/1000 - 1/10000. Detects a band of approximately 65, 70 kDa (predicted molecular weight: 65 kDa).
IHC-P 1/100.
ICC/IF 1/250.
Flow Cyt 1/40. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP 1/20 - 1/40.

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 (KO) 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 - ab182562 observed at 50-65 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab182562 was shown to specifically react with BIN1 when BIN1 knockout samples were used. Wild-type and BIN1 knockout samples were subjected to SDS-PAGE. Ab182562 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunoprecipitation. ab182562 at 1/10000 staining BIN1 in U87-MG cell lysate immunoprecipitated using ab182562 at 1/50.
    Lane 2: negative control.

  • Flow Cytometrical analysis of 2% paraformaldehyde fixed HeLa cells labeling BIN1 with ab182562 at 1/40.

  • Immunofluorescent analysis of HeLa cells labeling BIN1 with ab182562 at 1/250 and Goat anti rabbit IgG(Alexa Fluor®555) at 1/200.  Cells were fixed with -20℃ Acetone. Image at the right stained with DAPI.

  • Immunofluorescent analysis of U87-MG cells labeling BIN1 with ab182562 at 1/250 and Goat anti rabbit IgG(Alexa Fluor®555) at 1/200. Cells were fixed with 4% paraformaldehyde. Image at the right stained with DAPI.

  • Immunohistochemical analysis of paraffin embedded Human brain tissue labeling BIN1 with ab182562 at 1/100.

  • Immunohistochemical analysis of paraffin embedded Human clear cell carcinoma of kidney tissue labeling BIN1 with ab182562 at 1/100.

  • Immunohistochemical analysis of paraffin embedded Human skeletal muscle tissue labeling BIN1 with ab182562 at 1/100.

  • Anti-BIN1 antibody [EPR13463] (ab182562) at 1/20000 dilution + Human fetal brain at 20 µg

    Secondary
    Goat Anti-Rabbit IgG HRP (ab136636) at 1/500 dilution

    Predicted band size : 65 kDa
    Additional bands at : 90 kDa. We are unsure as to the identity of these extra bands.
  • All lanes : Anti-BIN1 antibody [EPR13463] (ab182562) at 1/5000 dilution

    Lane 1 : Human skeletal muscle
    Lane 2 : Human fetal kidney

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG HRP (ab136636) at 1/500 dilution

    Predicted band size : 65 kDa
    Additional bands at : .
  • All lanes : Anti-BIN1 antibody [EPR13463] (ab182562) at 1/5000 dilution

    Lane 1 : A431 cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size : 65 kDa
    Additional bands at : .

References

ab182562 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (iPS cell derived neurons)
Permeabilization
Yes - see blocking buffer
Specification
iPS cell derived neurons
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde
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Submitted Feb 02 2016

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