Biolegend, 421934, Cellular Senescence Kit Red
100tests
Catalog Number: 421934
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Category
Pack Size1 vial
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| Verified Reactivity | Mouse, Human |
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| 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 |
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| Application Notes |
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| 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 |
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| Function | Cellular Senescence activity |
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| Biology Area | Cell Biology, Cell Cycle/DNA Replication, Cell Proliferation and Viability |
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| Molecular Family | Enzymes and Regulators |
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| Gene ID | NA |
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| Regulatory Status | RUO |
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| Other Names | Senescence-associated beta-galactosidase |
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