Cellular Senescence Kit Red

Kit Contents

Kit Contents

• 1 vial of lyophilized 100X Senescence Probe Red
• 1 bottle (20 mL) of Senescence Assay Buffer
• 1 vial (100 µL) of DMSO

Product Details

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.

1. ​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.
2. 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:

1. 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).
2. Gently wash the cells with Cell Staining Buffer (Cat. No. 420201) or PBS.
3. 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.
4. Gently remove the Senescence Probe Red solution from cells and wash twice with Cell Staining Buffer (Cat. No. 420201) or PBS.
5. 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.

Antigen Details

Distribution

Cellular Senescence

Function

Cellular Senescence activity

Biology Area

Cell Biology, Cell Cycle/DNA Replication, Cell Proliferation and Viability

Molecular Family

Enzymes and Regulators

Gene ID

NA

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Pages & Pathways

Related FAQs

Certificate of Analysis