Description
Description: This assay employs a two-site sandwich ELISA to quantitate CYLC1 in samples. An antibody specific for CYLC1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CYLC1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CYLC1 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 CYLC1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Cylicin 1 contains numerous lysine dipeptides followed by a third variable amino acid (KKX), which in most cases is aspartic acid; these KKX tripeptides occur throughout the molecule, with the exception of the C-terminal tail, which contains proline-rich segments.The central portion of the protein is arranged as a series of repeating units that are predicted to form individual short alpha helices, which are interrupted by short linker segments. Antibodies against human cylicin-1 reacted with 2 polypeptides of 60 kD and 80 kD by immunoblot analysis and stained specifically the calyx of human and bovine spermatozoa by immunofluorescence microscopy. Northern blot analysis of bovine tissues detected a cylicin-1 transcript only in testis