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Product key features
- Sensitive detection: Colorimetric assay for easy visualization of senescent cells under a light microscope.
- Optimized reagents: Includes Xite™ beta-D-galactopyranoside substrate for accurate β-galactosidase activity detection.
- Simple protocol: Minimal hands-on time with easy-to-follow steps.
- Compatible with multiple cell types: Works with various adherent cell lines to study senescence-related processes.
Product description
The Cell Meter™ Colorimetric Cellular Senescence Activity Assay Kit is an easy-to-use method for detecting senescent cells in culture. Cellular senescence is a natural process in which actively dividing cells transition into a permanent, non-dividing state, often associated with aging. Senescence is accompanied by morphological changes, altered gene expression, and distinct physical characteristics. Over time, senescent cells typically progress toward cell death, although they can remain metabolically active for extended periods.
This kit utilizes the Xite™ beta-D-galactopyranoside substrate to enable the visualization of β-galactosidase activity, a key biomarker of cellular senescence, forming a with a distinct blue color as seen under a standard light microscope. It includes all necessary reagents for fixation, staining, and detection, ensuring high reproducibility. The simple workflow involves a straightforward incubation and washing process, allowing researchers to achieve reliable results without requiring fluorescence microscopy or specialized equipment. It is ideal for studying aging, cellular stress, and drug-induced senescence in cells.
Example protocol
AT A GLANCE
- Grow cells in a desired growth medium
- Remove the medium and fix cells with Fixation Solution
- Incubate the cells for 20 mins at RT
- Remove the Fixation Solution and wash cells twice with PBS
- Add Xite™ beta-D-galactopyranoside working solution
- Incubate cells for overnight at 37 ℃
- Monitor staining using light microscopy
Note: Before use, bring all the kit components at room temperature and centrifuge briefly to collect at the bottom.
PREPARATION OF STOCK SOLUTIONS
Xite™ beta-D-galactopyranoside stock solution (20X):
Prepare stock solution by adding 500 μL of DMSO (Component F) into Xite™ beta-D-galactopyranoside vial (Component E).
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
PREPARATION OF WORKING SOLUTION
To prepare 1 mL of Xite™ beta-D-galactopyranoside working solution, mix 930 μL of Staining Buffer (Component B), 10 μL of Additive Solution A (Component C), 10 μL of Additive Solution B (Component D) and 50 μL of Xite™ beta-D-galactopyranoside stock solution.
Note: For best results, this solution should be used within a few hours of its preparation.
Note: 10 mL of working solution is enough for 100 tests.
SAMPLE EXPERIMENTAL PROTOCOL
- Plate cells as desired in a 96-well black wall-clear bottom plate.
- Treat cells as desired to induce senescence.
- Remove the cell culture medium and wash cells with HHBS buffer.
- Add Fixation Solution and incubate cells for 20 minutes at RT.
- Remove the Fixation Solution and wash cells twice with PBS.
- Add 100 µL of Xite™ beta-D-galactopyranoside working solution to cells.
- Incubate cells at 37 º C for overnight, protected from light.
- Remove the dye working solution and wash cells twice with PBS.
- Add PBS and acquire images with light microscopy.
Note: The incubation must be done in 37ºC hot plate, since presence of CO2 in incubator can alter the pH of the solution.
References
Authors: Chen, Peng and Yang, Xian and Chen, Weijie and Wei, Wenwei and Chen, Yujie and Wang, Peiyuan and He, Hao and Liu, Shuoyan and Zheng, Yuzhen and Wang, Feng
Journal: Gene (2025): 149233
Authors: Wang, Gang and Wang, Yi and Xiao, Yanyi and Lin, Zhe
Journal: Scientific reports (2025): 5251
Authors: Deng, Guiyan and Xu, Cheng and Mo, Dunchang
Journal: PeerJ (2025): e18818
Authors: Wei, Chunyang and Liu, Xiaoying and Miao, Zhuang and Zhang, Hua and Wang, Yanfu and Qi, Guoxian
Journal: Future science OA (2025): 2455906
Authors: Shen, Zhiming and Gao, Yang and Sun, Xuedong and Chen, Min and Cen, Changhuo and Wang, Mengyue and Wang, Nan and Liu, Bowen and Li, Jiayi and Cui, Xiuhong and Hou, Jian and Shi, Yuhua and Gao, Fei
Journal: Cell proliferation (2025): e13760
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