Flow cytometric analysis of MCF7 cells, untreated (blue) or treated with Etoposide #2200 (12.5 μM, 24 hr; green) and allowed to recover for 3 days, using SA-β-Gal Fluorescent Substrate (solid lines) compared to unlabeled MCF7 cells (dashed line).
Additional Reagents (Not Supplied):
NOTE: Suggested dilutions and volumes are provided for reference. Optimal conditions for SA-beta-Gal Fluorescent Substrate performance may vary by cell type and container type.
NOTE: Cell treatments to induce senescence should be completed before initiating staining with this kit. Ensure that an untreated control is included, to provide a baseline measurement of fluorescence for comparison.
NOTE: Formaldehyde fixation may be performed at the completion of the incubation step with the SA-beta-Gal Fluorescent Substrate #38154. Fixation with up to 4% formaldehyde for up to 15 minutes will not significantly reduce the fluorescence of the substrate. However, permeabilization with methanol or Triton X-100 will cause a significant reduction in fluorescence. If permeabilization is required for immunolabeling of intracellular targets, conduct an experiment to determine whether this will significantly compromise detection of the fluorescent substrate in that cell model.
posted April 2010
Protocol Id: 1986
The Senescence β-Galactosidase Activity Assay Kit (Fluorescence, Flow Cytometry) allows the quantitative measure of senescence-associated β-galactosidase activity in living cells via flow cytometry using a cell permeable fluorogenic substrate, which becomes fluorescent upon hydrolysis by β-galactosidase. The fluorescent molecule is optimally excited at 490 nm (blue laser) and peak emission occurs at 514 nm (usually FL1). Each kit contains sufficient quantities of reagents to perform up to 100 assays.
Limited capacity to replicate is a defining characteristic of most normal cells and culminates in senescence, an arrested state in which the cell remains viable (1). Senescent cells are not stimulated to divide by serum or passage in culture, and senescence invokes a specific cell cycle profile that differs from most damage-induced arrest processes or contact inhibition (2). An enlarged cell size, expression of pH-dependent β-galactosidase activity (3), and an altered pattern of gene expression (4,5) further characterize senescent cells.
Explore pathways + proteins related to this product.
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.