Description
Description: This assay employs a two-site sandwich ELISA to quantitate LBH in samples. An antibody specific for LBH has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any LBH present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for LBH 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 LBH bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: The deduced 105-amino acid human and mouse proteins have a calculated molecular mass of 12 kD, and human LBH shares 90% amino acid identity with its mouse and bovine orthologs. The LBH proteins contain an N-terminal hydrophobic region, a putative nuclear localization signal, and a C-terminal acidic glutamate-rich domain. There are 2 predicted alpha helices that could act as protein-protein interaction interfaces. Immunohistochemical studies localized Lbh to the nucleus. Northern blot analysis of adult mouse tissues detected highest Lbh expression in the heart, followed by lung, spleen, brain, and kidney. Whole-mount in situ hybridization detected Lbh expression in embryonic bilateral heart primordia becoming more localized after heart tube fusion and looping, with highest expression in the right ventricle, atrioventricular canal, and sinus venosus