MarkerGene™ Fluorescent Cellular Senescence Microtiterplate Assay Kit
Product ID: M1405
Uses the high-sensitivity substrate fluorescein di-β-D-galactopyranoside (FDG) to quantify senescent cells.
Recent studies have shown that β-galactosidase histochemical staining at pH 6 is a useful enzymatic marker of cellular senescence (the so-called senescence associated beta-galactosidase; SA-β-Gal). While cells entering senescence show this enzymatic activity, immortalized cell types, including tumor or other quiescent cells, are not stained under these conditions. The implications of these methods in such diverse areas as age-related pathology research, tumor analysis and tissue culture maintenance is significant. FDG turnover produces the highly fluorescent product fluorescein, which is easily detected using an appropriate filter set for fluorescein fluorescence (i.e. EX 485nm and EM 535 nm) in a microtiterplate assay format. This assay kit contains all of the reagents for performing up to 100 assays using 6-well or 12-well tissue culture plate samples of cells or for cells that have been grown onto sterile cover slips for analysis from other plate sizes. In addition, the kit also contains reaction buffers, dilution solvents, a detailed protocol and references for use.
See also product Product M0250 for additional information and references.
|Unit Size||1 Kit|
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However, when we tested the kit using well characterized senescent cells obtained from collaborators, the staining patterns for X-Gal and FDG (SA-b-Gal) were very similar (see the data contained in the product information sheet provided in the kit).
We searched for other methods to provide a positive control (inducer of senescence) reagent to add to the kit, but without success. However, we feel confident that the FDG method can measure SA-bGal levels in senescent cells according to the methods described.
To date we have not been able to find a suitable positive control (reagent) to artificially induce senescence in cells lines for the kit, and therefore suggest verifying staining patterns by separate X-Gal staining.