Biolegend, 421934, Cellular Senescence Kit Red, 100tests

Cellular senescence is a state of stable cell cycle arrest under which cells remain metabolically active, but no longer divide and do not respond to growth-promoting stimuli. It is triggered by a variety of cellular stressors, including environmental factors like ionizing radiation or exposure to chemotherapeutic drugs, oxidative stress, DNA damage, mitochondrial dysfunction, and oncogene activation. Senescence-Associated β-galactosidase (SA-β-gal), a result of altered lysosomal activity, is a hallmark of cellular senescence. Cellular Senescence Kit Red uses a fluorogenic beta-Gal substrate that readily enters into live cells, where it gets cleaved by SA-β-gal inside cells, generating strong red fluorescence. It provides higher sensitivity compared to X-gal staining, a colorimetric method widely available and used to detect Senescence-associated beta-galactosidase (SA-beta-gal) in senescent cells.
100tests
Verified Reactivity: Mouse, Human
Formulation: Each kit contains 1 vial of lyophilized 100X Senescence Probe Red, 1 bottle (20 mL) of Senescence Assay Buffer, 1 vial (100 µL) of DMSO.
Storage & Handling: The kit should be stored at < -15°C. All unused stock solutions should be divided into single-use aliquots and stored at < -15°C after preparation. Avoid repeated freeze-thaw cycles.
Application: FC - Quality tested Live cell imaging - Verified
Recommended Usage: Please refer to detailed protocol in application notes
Application Notes: Components: 1 vial of lyophilized 100X Senescence Probe Red (beta-D-galactopyranoside) (Store at < -15°C and minimize light exposure. All unused stock solutions should be stored as single-use aliquots and stored at < -15°C. Avoid repeated freeze-thaw cycles.) 1 bottle (20 mL) of Senescence Assay Buffer (Store at < -15°C) 1 vial (100 μL) of DMSO (Store at < -15°C) Required Materials Not Included: Buffer: PBS Detection/Imaging Guidelines: Fluorescence microscope filter set: Cyanine3/TRITC filter set, recommend black wall/clear bottom plate Flow cytometry channel: Excitation: 488 nm laser Emission: 575/26 nm filter Instrument specification(s): PE channel Assay Protocol: ​This kit provides enough reagents for 100 tests. A test is defined as 100 µL of Senescence Probe Red working solution for up to 1x106 cells. ​Preparation of stock solutions: Add 100 µL of DMSO into lyophilized 100X Senescence Probe Red and mix well. Note: Store the unused Senescence Probe Red stock solution at -20°C in single use aliquots. Preparation of working solutions: Dilute 10 µL of 100X Senescence Probe Red stock solution with 1 mL of Senescence Assay Buffer to make Senescence Probe Red working solution. This can be used for 10 tests; each test will need 100 µL Senescence Probe Red working solution. Note: Senescence Probe Red working solution should be used promptly. Experimental Protocol: Treat your cell samples under desired experimental conditions. Note: For selectively tracking β-Gal in live cells, cells can be treated with Bafilomycin A1 for blocking endogenous β-Gal. Optimum concentration of Bafilomycin A1 may vary depending on cell type (suggested range is 100 nM-10 mM). Gently wash the cells with Cell Staining Buffer (Cat. No. 420201) or PBS. Add 100 µL Senescence Probe Red working solution for up to 1x106 cells (for higher cell amounts, scale up accordingly). And incubate the samples at 37°C incubator for 15-45 minutes. Gently remove the Senescence Probe Red solution from cells and wash twice with Cell Staining Buffer (Cat. No. 420201) or PBS. Analyze samples: For flow cytometry, resuspend the cells in the Senescence Assay Buffer and monitor the fluorescence intensity on a flow cytometer using a 575/26 nm filter (PE channel). For live cell imaging, observe cells with inverted fluorescence microscope using a Cyanine3/TRITC filter set.
Distribution: Cytoplasmic
Function: Cellular Senescence activity
Biology Area: Cell Biology, Cell Cycle/DNA Replication, Cell Proliferation and Viability
Molecular Family: Enzymes and Regulators
Gene ID: NA
Regulatory Status: RUO
Other Names: Senescence-associated beta-galactosidase