Description
Description: This assay employs a two-site sandwich ELISA to quantitate GAGE12G in samples. An antibody specific for GAGE12G has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any GAGE12G present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for GAGE12G 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 GAGE12G bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: GAGE12G belongs to a multigene family expressed in a large variety of tumors whereas in normal tissues, expression is restricted to germ cells. These genes organized in clustered repeats, have a high degree of predicted sequence identity, but differ by scattered single nucleotide substitution. Their sequences contain either the antigenic peptide YYWPRPRRY or YRPRPRRY which is recognized by cytotoxic T cells.
The first GAGE nomenclature was based on identified mRNA sequences, but the high identity of the GAGE members made impossible to separate products of paralogous genes from polymorph products. Ref.3 presented a new GAGE gene nomenclature based on the identified genes and their products