Quantitative Cellular Senescence Assay

Background/Introduction: Normal primary cells proliferate in culture for a limited number of population doublings prior to undergoing terminal growth arrest and acquiring a senescent phenotype. This finite life span correlates with the age of the organism and with the life expectancy of the species from which the cells were obtained; such that the older the age or the shorter the life span, the less the ability of the cells to undergo population doubling. Senescent cells are characterized by an irreversible G1 growth arrest involving the repression of genes that drive cell cycle progression and the upregulation of cell cycle inhibitors like p16INK4a, p53, and its transcriptional target, p21CIP1. They are resistant to mitogen-induced proliferation, and assume a characteristic enlarged, flattened morphology. Research into the pathways that positively regulate senescence and ways cells bypass senescence is therefore critical in understanding carcinogenesis. Normal cells have several mechanisms in place to protect against uncontrolled proliferation and tumorigenesis. Senescent cells show common biochemical markers such as expression of an acidic senescence-associated beta-galactosidase (SA-beta-Gal) activity. While senescence has been characterized primarily in cultured cells, there is also evidence that it occurs in vivo. Cells expressing markers of senescence such as SA-beta-Gal have been identified in normal tissues. Our Quantitative Cellular Senescence Assay Kit uses a fluorogenic substrate to measure SA-beta-Gal activity. This compound is a membrane permeable, non-fluorescent substrate of beta-galactosidase, which after hydrolysis of the galactosyl residues emits green fluorescence and remains confined within the cell. In comparison to the conventional cytochemical assay using X-gal, the use of a fluorogenic substrate greatly enhanced the sensitivity of the assay. Also, the non-toxic fluorogenic substrate allows you to study living cells by flow cytometry for more quantitative measurement of SA-beta-Gal activity. Each kit provides sufficient quantities to perform up to 10 assays in 35 mm wells.

1. Current Protocols in Molecular Biology, John Wiley & Sons Press.
2. Campisi, J. (2000) In Vivo 14, 183-188.
3. Dimri, G. P., X. Lee, G. Basile, M. Acosta, G. Scott, C. Roskelley, E. E. Medrano, M. Linskens, I. Rubelj, O. Pereira-Smith, M. Peacocke, and J. Campisi. (1995) Proc. Natl. Acad. Sci. USA 92:9363-9367.
1. Kim, J. et al. (2014) p53 Induces Skin Aging by Depleting Blimp1+ Sebaceous Gland Cells. Cell Death Dis. 5: e1141 2. Grasso, D. et al. (2014). Genetic Inactivation of the Pancreatitis-Inducible Gene Nupr1 Impairs PanIN Formation by Modulating Kras(G12D)-Induced Senescence. Cell Death Differ. 21:1633-1641. 3. Landowski, T. H. et al. (2014). Targeting Integrin alpha6 Stimulates Curative-Type Bone Metastasis Lesions in a Xenograft Model. Mol Cancer Ther. 13:1558-1566.

Small volumes of Quantitative Cellular Senescence Assay assay kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS169055 is an assay kit targeting Quantitative Cellular Senescence Assay.