The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at 5-15 µg/mg of lysate.
1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml. Predicted molecular weight: 119 kDa. This antibody has not performed satisfactorily when used for WB of Importin4 in crude preparations (e.g. whole cell lysate). ab99270 can be used for WB of enriched (e.g. immunoprecipitated) sources of Importin4.
Functions in nuclear protein import as nuclear transport receptor. Serves as receptor for nuclear localization signals (NLS) in cargo substrates. Is thought to mediate docking of the importin/substrate complex to the nuclear pore complex (NPC) through binding to nucleoporin and the complex is subsequently translocated through the pore by an energy requiring, Ran-dependent mechanism. At the nucleoplasmic side of the NPC, Ran binds to the importin, the importin/substrate complex dissociates and importin is re-exported from the nucleus to the cytoplasm where GTP hydrolysis releases Ran. The directionality of nuclear import is thought to be conferred by an asymmetric distribution of the GTP- and GDP-bound forms of Ran between the cytoplasm and nucleus (By similarity). Mediates the nuclear import of RPS3A. In vitro, mediates the nuclear import of human cytomegalovirus UL84 by recognizing a non-classical NLS.
Belongs to the importin beta family. Contains 6 HEAT repeats. Contains 1 importin N-terminal domain.
Detection of Importin4 in Immunoprecipitates of HeLa whole cell lysate (1 mg for IP, 20% of IP loaded) using ab99270 at 10 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection. Lane 2 represents control IgG IP. Detection: Chemiluminescence with an exposure time of 5 seconds