Overview

  • Product name
  • Description
    Mouse monoclonal to BIN1
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment: VVETFPATVN GTVEGGSGAG RLDLPPGFMF KVQAQHDYTA TDTDELQLKA GDVVLVIPFQ NPEEQDEGWL MGVKESDWNQ HKELEKCRGV FPENFTERVP , corresponding to amino acids 355-455 of Human BIN1

  • Positive control
    • ICC/IF: SKNSH cells.
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause.

Properties

Applications

Our Abpromise guarantee covers the use of ab54764 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 Use a concentration of 1 - 5 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 10 µg/ml.

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

  • 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 - ab54764 observed at 45-65 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab54764 was shown to specifically react with BIN1 when BIN1 knockout samples were used. Wild-type and BIN1 knockout samples were subjected to SDS-PAGE. Ab54764 and ab181602 (Rabbit 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • ICC/IF image of ab54764 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab54764, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing SH-SY5Y cells stained with ab54764 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab54764, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • BIN1 antibody (ab54764) at 1ug/lane + HeLa cell lysate at 25ug/lane.

References

ab54764 has not yet been referenced specifically in any publications.

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

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