The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
1/1000 - 1/10000. Detects a band of approximately 122 kDa (predicted molecular weight: 122 kDa).
Myosins are actin-based motor molecules with ATPase activity. Unconventional myosins serve in intracellular movements. Their highly divergent tails are presumed to bind to membranous compartments, which would be moved relative to actin filaments. Involved in glucose transporter recycling in response to insulin by regulating movement of intracellular GLUT4-containing vesicles to the plasma membrane. Component of the hair cell's (the sensory cells of the inner ear) adaptation-motor complex. Acts as a mediator of adaptation of mechanoelectrical transduction in stereocilia of vestibular hair cells. Binds phosphoinositides and links the actin cytoskeleton to cellular membranes. Isoform 3 is involved in regulation of transcription. Associated with transcriptional active ribosomal genes. Appears to cooperate with the WICH chromatin-remodeling complex to facilitate transcription. Necessary for the formation of the first phosphodiester bond during transcription initiation.
Contains 2 IQ domains. Contains 1 myosin head-like domain.
Binds directly to large unilamellar vesicles (LUVs) containing phosphatidylinositol 4,5-bisphosphate (PIP2) or inositol 1,4,5-trisphosphate (InsP3). The PIP2-binding site corresponds to a putative PH domain present in its tail domain.
Cytoplasm. Cell membrane. Cell projection > stereocilium membrane. Colocalizes with CABP1 and CIB1 at cell margin, membrane ruffles and punctate regions on the cell membrane. Colocalizes in adipocytes with GLUT4 in actin-based membranes. Localizes transiently at cell membrane to region known to be enriched in PIP2. Activation of phospholipase C results in its redistribution to the cytoplasm and Nucleus > nucleoplasm. Nucleus > nucleolus. Nucleus > nuclear pore complex. Colocalizes with RNA polymerase II in the nucleus. Colocalizes with RNA polymerase I in nucleoli (By similarity). In the nucleolus, is localized predominantly in dense fibrillar component (DFC) and in granular component (GC). Accumulates strongly in DFC and GC during activation of transcription. Colocalizes with transcription sites. Colocalizes in the granular cortex at the periphery of the nucleolus with RPS6. Colocalizes in nucleoplasm with RPS6 and actin that are in contact with RNP particles. Colocalizes with RPS6 at the nuclear pore level.
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Myosin 1C with ab194828 at 1/200 dilution (5μg/ml). A Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution was used as secondary (ab97051). Counterstain: Hematoxylin. Inset image: negative control obtained using PBS instead of ab194828. Cytoplasm staining on Human tonsil was observed.
Immunofluorescent analysis of A431 cells labeling Myosin 1C with ab194828 at 1/100 dilution. A Goat anti rabbit IgG (Alexa Fluor488) at 1/400 dilution (ab150077) was used as secondary antibody. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% triton X-100. Counterstain: DAPI. Cytoplasm and nuclear on A431 cell line was observed.
Western blot - Anti-Myosin 1C antibody [EPR14771] (ab194828)
All lanes : Anti-Myosin 1C antibody [EPR14771] (ab194828) at 1/2000 dilution
Lane 1 : Mouse heart lysate Lane 2 : Rat heart lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Flow cytometry analysis of HeLa cells labelling Myosin 1C (red) with purified ab194828 at dilution of 1/80. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.