Components

Fixative Solution, X-Gal, Staining Solution, Staining Supplement, and a user protocol.

General description

Useful for the histochemical detection of senescent cells. The failure of cells to become senescent may lead to tumor progression. Higher levels of β-Galactosidase is considered as a good marker for senescent cells. The mechanism of this is believed to be due to the increased lysosomal content of senescent cells, resulting in an increase of the lysosomal enzyme β-galactosidase. Cells can be observed using microscope.

Senescence is thought to be a tumor suppressive mechanism and an underlying cause of aging. It represents an arrested state in which the cells remain viable, but not stimulated to divide by serum or by passage in culture. Senescent cells display an increase in cell size, pH-dependent expression of β-galactosidase (β-Gal) activity, and altered patterns of gene expression.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Kurz, D.J., et al. 2000. J. Cell Sci.113, 3613.Dimri, G.P., et al. 1995. Proc. Natl. Acad. Sci. USA92, 9363.

Packaging

1 kit in Glass bottle

Preparation Note

PBS: Prepare a 1X PBS solution (not provided). Use at least 3 ml per well. X-gal: Dissolve 20 mg X-gal in 1 ml of DMF (N-N-dimethylformamide, not provided) to prepare a 20X stock solution. Aliquot and freeze (-20°) the unused X-gal solution. This stock solution is stable for up to 1 month at -20°C (protect from light). Always store X-gal in a polypropylene or glass container. Do not use polystyrene. Staining Solution Mix: While cells are incubating in Fixative Solution (step 2 below), prepare Staining Solution Mix in a polypropylene plastic tube by adding 470 µl of Staining Solution, 5 µl Staining Supplement, and 25 µl of 20 mg/ml X-gal in DMF.

Principle

The Calbiochem^® Senescence Detection Kit is useful for the histochemical detection of senescent cells. The failure of cells to become senescent may lead to tumor progression. The mechanism of this is believed to be the increased lysosomal content of senescent cells resulting in an increase of the lysosomal enzyme β-galactosidase.

Specifications

Assay Time: 30 min. plus overnight incubation

Storage and Stability

Upon arrival store the entire contents of the kit at -20°C.

Warning

Toxicity: Multiple Toxicity Values, refer to MSDS (O)