Description
Description: This assay employs a two-site sandwich ELISA to quantitate FSCN3 in samples. An antibody specific for FSCN3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any FSCN3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for FSCN3 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 FSCN3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Mapping of murine and human FSCN3 genes shows localization to the 7q31.3 chromosome. FSCN3 expression is highly specific to testis and that in situ hybridization further restricts expression to elongating spermatids. Antibodies raised against recombinant FSCN3 protein identify a band at 56 kD in testis, epididymis, and epididymal spermatozoa, suggesting that testis fascin persists in mature spermatozoa. In accord with the in situ hybridization results, immunofluorescent microscopy localizes testis fascin protein to areas of the anterior spermatid head that match known distributions of F-actin in the dorsal and ventral subacrosomal spaces. It is possible that testis fascin may function in the terminal elongation of the spermatid head and in microfilament rearrangements that accompany fertilization