cell death drug screening in cancer models apoptosis necrosis ELISA assay test kit

Peviva Products are for Research Use Only

Three steps to oncology drug research discovery:

  1. Start with cell culture work to screen for apoptosis-inducing cancer compounds
  2. Test effect of compounds in animal models (human tumor xenograft models)
  3. Measure response to cancer compound using serum or plasma samples

The M30 CytoDeath™ ELISA is for apoptosis measurement in cell culture applications. The assay has a dynamic range and sensitivity suitable for in vitro characterization of apoptosis-inducing drugs, including establishment of time course kinetics and dose-response relationships. The effects of large numbers of compounds on apoptosis can be tested in the 96-well format and the assay can quantify apoptosis of epithelially derived cells in multicellular spheroids and tissue culture. A additional key research  feature is that the CK18 kits measure an accumulated caspase-derived product, so there is no risk to miss time of apoptosis and transient caspase activation.

The release of intracellular proteins from dying tumour cells is possible to exploit as a PD marker for response to cytotoxic anticancer research. One type of intracellular protein commonly used for this purpose are the cytokeratins (CKs), which are highly expressed in carcinomas but not by chemotherapy-sensitive cells of the bone marrow. Lung, breast, prostate and colon carcinomas are the cancermost frequent forms of malignancies. These tumors all arise from simple epithelial cells and express the intermediate filament protein cytokeratin 18 (CK18, keratin 18 or K18). During apoptosis, CK18 is cleaved by caspases at two sites. Macrophages recognize and engulph apoptotic cells. This does not necessarily occur in tumor tissue and caspase-cleaved CK18 may therefore be released to the extracellular environment and into the blood. Apoptosis of other cell types such as lymphoid cells will not generate CK18 cleavage products. Caspase-cleaved CK18 is therefore an ideal biomarker for measuring apoptosis of carcinoma cells using blood samples in research.

The M30 Apoptosense CK18 research kit specifically detects caspase-cleaved CK18 (ccCK18) released from apoptotic tumor cells. The M30 Apoptosense® ELISA assay is designed to be used together with the M65 ELISA which measures total CK18.  Determination of the ratio between caspase-cleaved to total CK18 released from dead cells offers a unique possibility to determine cell death mode (apoptosis or necrosis).

Tumor cells die in vivo by apoptosis and necrosis. The mode of cell death depends both on the type of cytotoxic stimulus and the tumor. An apoptotic stimulus may under conditions of deficient cellular ATP generation fail to induce the apoptotic program and cells instead undergo necrosis. During necrosis, intracellular proteins that are not cleaved by caspases will be released from cells.

Many anticancer agents induce apoptosis. Apoptosis is also an attractive research endpoint when studying novel therapeutic drugs in oncology. The M30 Apoptosense CK18 and M65 CK18 kit are unique assays for studying tumor cell death during cancer research. Effective cancer compounds induce increases in caspase-cleaved and/or total CK18 in the circulation within 24-72 hours.

 

cell death drug screening in cancer models apoptosis necrosis ELISA assay test kit

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