The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/1000. Detects a band of approximately 56, 65, 70, 90 kDa (predicted molecular weight: 65 kDa).
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 [EPR13463-25] (ab185950)
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).