详细信息:ssDNA Apoptosis ELISA Kit.This procedure is based on the selective denaturation of DNA in apoptotic cells by formamide, and detection of denatured DNA with monoclonal antibody to single-stranded DNA (ssDNA). Formamide is a gentle agent that denatures DNA in apoptotic cells, but not in necrotic cells or in the cells with DNA breaks in the absence of apoptosis (Frankfurt & Krishan 2001). The sensitivity of DNA in apoptotic cells to formamide is not related to DNA breaks, but rather reflects changes in chromatin associated with apoptosis, such as chromatin condensation and digestion of proteins stabilizing DNA. The assay includes attachment of cells to 96-well plates, treatment of attached cells with formamide, and staining of ssDNA in apoptotic cells with a mixture of primary antibody and peroxidase-labeled secondary antibody. The protocol based on the one-step detection of ssDNA with antibody mixture has higher sensitivity and lower number of steps than standard two-step immunostaining (Frankfurt & Krishan 2001). This mixture is included in the kit in a ready to use form.
