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.
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
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/10000. Detects a band of approximately 65, 70 kDa (predicted molecular weight: 65 kDa).
1/40. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/20 - 1/40.
May be involved in regulation of synaptic vesicle endocytosis. May act as a tumor suppressor and inhibits malignant cell transformation.
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.
Western blot - Anti-BIN1 antibody [EPR13463] (ab182562)
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.
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.