Description
Description: This assay employs a two-site sandwich ELISA to quantitate IPO5 in samples. An antibody specific for IPO5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IPO5 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for IPO5 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IPO5 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: The import of proteins containing a nuclear localization signal (NLS) requires the NLS import receptor, a heterodimer of importin alpha and beta subunits also known as karyopherins. Importin alpha binds the NLS-containing cargo in the cytoplasm and importin beta docks the complex at the cytoplasmic side of the nuclear pore complex. In the presence of nucleoside triphosphates and the small GTP binding protein Ran, the complex moves into the nuclear pore complex and the importin subunits dissociate. Importin alpha enters the nucleoplasm with its passenger protein and importin beta remains at the pore. Interactions between importin beta and the FG repeats of nucleoporins are essential in translocation through the pore complex. Importin 5 is a member of the importin beta family