MB49 In Vitro Caspase Activation-based Apoptosis Assay (FCM)

This assay is to provide MB49-based In Vitro Caspase Activation-based Apoptosis Assay (FCM) to accelerate our client's oncology projects. The assay will be customized according to the specific requirements. Please contact our scientists to discuss more details.

MB49

Mouse

MB49 cells are derived from C57BL/Icrf-a' mouse bladder epithelial cells that were transformed by a single 24-hour treatment with the chemical carcinogen 7, 12-dimethylbenz[a]anthracene (DMBA) on the second day of a long term primary culture. Transformed cells transplanted into syngeneic mice were shown to generate carcinomas. While of male origin, karyotype analyses indicate the loss of the Y chromosome in 100% of the cells analyzed. This abnormality is a frequent early event in human bladder cancer.

Bladder Cancer

Oncology

In Vitro Caspase Activation-based Apoptosis Assay (FCM)

MB49-cell based In Vitro Caspase Activation-based Apoptosis Assay (FCM)

Activation of caspases in cancer cells can also be detected by FCM using caspase-specific short peptides linked with a fluorophore. For example, when rhodamine 110 (green fluorescent dye)-linked caspase-specific peptide is attached to protein caspase, its fluorescence is quenched and can be detected by FCM. Fluorochrome-labeled inhibitor of caspase assay (FLICA) is another method used for the detection of apoptosis. Here, fluorylmethyl ketone and carboxyfluorescein are attached at both the ends of a peptide specific to caspase. Upon binding with caspase proteins, it is cleaved and fluoryl methyl ketone prevents further activity by binding with its active sites and finally fluorescence signals will build up in cells.

Detection of Apoptosis Assays

Apoptosis (programmed cell death) plays a vital role in embryonic development, homeostasis, functioning of immune system and wound repair. The ability to evade induction of apoptosis has been used by cancer cells to survive against host defense mechanisms. The molecular mechanisms involved in cancer cell apoptosis have been well documented and it involves certain biochemical events such as DNA fragmentation, chromatin condensation, cell organelle degradation and protein cleavage, etc. The extrinsic and intrinsic (mitochondrial) pathways are the two major pathways involved in apoptosis. With the available techniques and assays, a number of apoptosis inducing agents (natural compounds, synthetic compounds, nano-formulations, peptides and enzymes) in many cancer cells have been identified. Selection of an assay for apoptosis detection is based on factors such as apoptotic pathway, nature of drug, cell type being used and the method of analysis.

Flow Cytometry to Detect Apoptosis Assays

Flow Cytometry Assay