Cellular senescence is a phenomenon by which normal cells cease to divide and enter a state of irreversible growth arrest. It is one of the most fundamental aspects of cellular behavior and generally believed to be associated with tumor suppressive mechanisms and an underlying cause of aging. Senescence-associated ;-galactosidase (SA-;-gal), which is overexpressed in senescent cells, has been widely used as a marker of cellular senescence. Although X-gal is a well-known reagent used to detect SA-;-gal, X-gal has the following disadvantages: 1) requirement of fixed cells due to poor cell-permeability, 2) low quantitative capability because of the difficulty in visually differentiating stained vs. non-stained cells 3) requirement for long stain times.
To conquer the disadvantages inherent in using X-gal, we utilize a new reagent and method for the detection of cellular senescence (Figure 1). SPiDER-;Gal offers increased sensitivity in detecting SA-;-gal due to increased cell permeability and retention inside cells. Compared to the X-gal method, the FastCellular Senescence Detection Kit can be used for both living and fixed cells, because we provide Bafilomycin A1 to specifically inhibit endogenous ;-galactosidase activity in living cells and thus increase the detection specificity for living cells. For fixed cells, SA-;-gal is detectable by using SPiDER-;Gal and Mcllvaine buffer (pH 6.0, please refer to the protocol for fixed cell assays). Since SPiDER-;Gal emits strong and stable fluorescence after the reaction with SA-;-gal, it can be applied to quantitative analysis by flow cytometry. In addition, the staining time is only 30 mins. Kit includes a 6-well plate with 1 unit of SPiDER-;Gal and 1 unit of Bafilomycin A1
Benefits of using FastCellular Senescence Detection Kit:
- Higher sensitivity due to a new fluorogenic detection probe
- Applicable for living cells and fixed tissues
- Ability to quantify SA-;-gal in cellular senescence assay
- Short staining time (30 min)
1 Kit