DiaPharma Group, Inc.

M30-Apoptosense ELISA is a unique tool for reliable and reproducible quantitative detection of apoptosis of CK18-positive cells.

The assay is applicable to in vitro cultured cells, tumor extracts and patient sera. All reagents are provided in a convenient ready-for-use format. The separable strips provide the flexibility for your experimental design!

M30-Apoptosense is an enzyme-linked immunosorbent assay (ELISA) for the quantitative detection of caspase-cleaved cytokeratin-18 (CK18Asp396-NE: M30 neo-epitope) levels in cytosolic cell extracts or the cell culture supernatants, human serum or plasma or other body fluids. The M30-Apoptosense ELISA uses the M30 antibody which is specific to this CK18 cleavage site and does not react with viable or necrotic cells (Leers et al., 1999).

M30-Apoptosense ELISA measures the accumulation of a caspase-generated product resulting in an unmatched sensitivity thus allowing the tracing of apoptotic cells of epithelial origin (e.g. normal epithelial cells or carcinoma cells) in patient sera (Ueno et al., 2003; Kramer et al., 2004; Roth et al., 2004). Due to this principle, the assay is ideally suited for high through-put screening as well (Hägg et al. 2002; Erdal et al. 2005).

Background

The most frequent forms of cancer in humans arise from cells of simple epithelia and express cytokeratin-18 (CK18). The unique combination of high expression in most cancer cells, but not in non-epithelial (lymphoid) cells, and the apoptosis specific cleavage by caspases makes CK18 an ideal marker for monitoring cancer cell apoptosis in human sera. Recent studies have indeed showed increased levels of caspase-cleaved CK18 (CK18Asp396-NE: M30 neo-epitope) in sera from cancer patients subjected to different treatment regimen (Ueno et al., 2003; Bivén et al. 2003; Kramer et al., 2004)

M30-Apoptosense is an ELISA based on the M30 monoclonal antibody (Leers et al., 1999) and therefore can be used for the quantitative detection of the M30 neo-epitope (CK18Asp396-NE) on CK18. As the M30 neo-epitope is only exposed after caspase-cleavage of CK18 (Hägg et al. 2002), it represents a specific marker of apoptosis in epithelial cells.

As the principle of the M30-Apoptosense ELISA is to measure the accumulation of a caspase cleavage product rather than proteolytic activity itself, it represents an advantageous assay system for high through-put screening (HTS) projects: it avoids the necessity to analyse multiple time-points after the apoptotic stimulus making it cost-effective as well (Hägg et al. 2002; Bivén et al. 2003; Hägg et al. 2004; Erdal et al. 2005).