Description
Description: This assay employs a two-site sandwich ELISA to quantitate POLI in samples. An antibody specific for POLI has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any POLI present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for POLI 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 POLI bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: The structure reveals a polymerase that is specialized for Hoogsteen basepairing, whereby the templating base is driven by to syn conformation. Mouse and human RAD30B proteins share approximately 74% amino acid identity. Members of the RAD30-like family of proteins share 5 conserved motifs clustered in their respective N termini. The C termini are much more diverged and vary considerably in length. Northern blot analysis detected an approximately 3-kb transcript in all human tissues tested, with elevated expression in testis. Expression in heart and pancreas was detected to a lesser extent than in testis, and low levels were present in other tissues. In situ hybridization analysis indicated that expression of mouse Rad30b occurs predominantly in postmeiotic round spermatids