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Xite™ Green beta-D-galactopyranoside
Xite™ Green beta-D-galactopyranoside is a fluorogenic substrate for beta-galactosidase (β-gal). Compared to the existing beta-galactosidase substrates (e.g., the commonly used FDG), it has much better cell permeability. Xite™ Green beta-D-galactopyranoside readily enters cells where it gets cleaved by β-gal, producing Xite™ Green, a strongly fluorescent product. The strongly fluorescent Xite™ Green is well retained in cells, making it easy to be detected with a flow cytometer and fluorescence microscope. Xite™ Green beta-D-galactopyranoside provides a simple and sensitive tool to detect beta-galactosidase activity. Xite™ Green beta-D-galactopyranoside might be used as a simple tool for measuring cellular senescence in cells since β-gal has been identified as a reliable marker for cellular senescence.
Example protocol
AT A GLANCE
Protocol summary
- Treat samples as desired
- Prepare and add Xite™ Green beta-D-galactopyranoside working solution to samples
- Incubate samples at 37 °C for 15 to 45 minutes
- Monitor the fluorescence intensity using flow cytometer with 530/30 nm filter (FITC channel) or using fluorescence microscopy with FITC filter set
PREPARATION OF STOCK SOLUTIONS
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.
Xite™ Green beta-D-galactopyranoside stock solution
Add appropriate amount of DMSO into Xite™ Green beta-D-galactopyranoside to make 2-5 mM Xite™ Green beta-D-galactopyranoside stock solution. Note: Store the unused Xite™ Green beta-D-galactopyranoside stock solution at -20 °C in single use aliquots.PREPARATION OF WORKING SOLUTION
Xite™ Green beta-D-galactopyranoside working solution
Prepare 1-20 µM of Xite™ Green beta-D-galactopyranoside working solution in buffer of your choice. Note: Xite™ Green beta-D-galactopyranoside working solution should be used promptly. Note: The concentration of the Xite™ Green beta-D-galactopyranoside should be optimized for different cell types and conditions.SAMPLE EXPERIMENTAL PROTOCOL
The following protocol can be used as a guideline and should be optimized according to the needs.
- Treat your samples as desired.
- Remove the treatment and wash the cells with buffer of your choice such as DPBS. 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 on type of cells.
- Add Xite™ Green beta-D-galactopyranoside working solution for 15-45 minutes and incubate the samples at 37 °C incubator. Note: Optimal time for incubation needs to be determined experimentally.
- Remove the working solution and wash cells with buffer of your choice.
- Resuspend the cells in buffer of your choice and monitor the fluorescence intensity with flow cytometer using 530/30 nm filter (FITC channel) or fluorescence microscope with FITC filter set.
Citations
View all 1 citations: Citation Explorer
Inhibiting the Histone Demethylase Kdm4a Restrains Cardiac Fibrosis After Myocardial Infarction by Promoting Autophagy in Premature Senescent Fibroblasts
Authors: Jin, Ming and Li, Chuling and Wu, Zhaoyi and Tang, Zhenquan and Xie, Jingfang and Wei, Guoquan and Yang, Zhiwen and Huang, Senlin and Chen, Yijin and Li, Xinzhong and others,
Journal: Advanced Science (2025): 2414830
Authors: Jin, Ming and Li, Chuling and Wu, Zhaoyi and Tang, Zhenquan and Xie, Jingfang and Wei, Guoquan and Yang, Zhiwen and Huang, Senlin and Chen, Yijin and Li, Xinzhong and others,
Journal: Advanced Science (2025): 2414830
References
View all 18 references: Citation Explorer
Novel fluorescent probe for rapid and ratiometric detection of β-galactosidase and live cell imaging.
Authors: Chen, Xiangzhu and Zhang, Xueyan and Ma, Xiaodong and Zhang, Yuanyuan and Gao, Gui and Liu, Jingjing and Hou, Shicong
Journal: Talanta (2019): 308-313
Authors: Chen, Xiangzhu and Zhang, Xueyan and Ma, Xiaodong and Zhang, Yuanyuan and Gao, Gui and Liu, Jingjing and Hou, Shicong
Journal: Talanta (2019): 308-313
Targeting senescence improves angiogenic potential of adipose-derived mesenchymal stem cells in patients with preeclampsia.
Authors: Suvakov, Sonja and Cubro, Hajrunisa and White, Wendy M and Butler Tobah, Yvonne S and Weissgerber, Tracey L and Jordan, Kyra L and Zhu, Xiang Y and Woollard, John R and Chebib, Fouad T and Milic, Natasa M and Grande, Joseph P and Xu, Ming and Tchkonia, Tamara and Kirkland, James L and Lerman, Lilach O and Garovic, Vesna D
Journal: Biology of sex differences (2019): 49
Authors: Suvakov, Sonja and Cubro, Hajrunisa and White, Wendy M and Butler Tobah, Yvonne S and Weissgerber, Tracey L and Jordan, Kyra L and Zhu, Xiang Y and Woollard, John R and Chebib, Fouad T and Milic, Natasa M and Grande, Joseph P and Xu, Ming and Tchkonia, Tamara and Kirkland, James L and Lerman, Lilach O and Garovic, Vesna D
Journal: Biology of sex differences (2019): 49
SA-β-Galactosidase-Based Screening Assay for the Identification of Senotherapeutic Drugs.
Authors: Fuhrmann-Stroissnigg, Heike and Santiago, Fernando E and Grassi, Diego and Ling, YuanYuan and Niedernhofer, Laura J and Robbins, Paul D
Journal: Journal of visualized experiments : JoVE (2019)
Authors: Fuhrmann-Stroissnigg, Heike and Santiago, Fernando E and Grassi, Diego and Ling, YuanYuan and Niedernhofer, Laura J and Robbins, Paul D
Journal: Journal of visualized experiments : JoVE (2019)
Cellular and cytoskeletal alterations of scleral fibroblasts in response to glucocorticoid steroids.
Authors: Bogarin, Thania and Saraswathy, Sindhu and Akiyama, Goichi and Xie, Xiaobin and Weinreb, Robert N and Zheng, Jie and Huang, Alex S
Journal: Experimental eye research (2019): 107774
Authors: Bogarin, Thania and Saraswathy, Sindhu and Akiyama, Goichi and Xie, Xiaobin and Weinreb, Robert N and Zheng, Jie and Huang, Alex S
Journal: Experimental eye research (2019): 107774
Tumor cell escape from therapy-induced senescence.
Authors: Saleh, Tareq and Tyutyunyk-Massey, Liliya and Murray, Graeme F and Alotaibi, Moureq R and Kawale, Ajinkya S and Elsayed, Zeinab and Henderson, Scott C and Yakovlev, Vasily and Elmore, Lynne W and Toor, Amir and Harada, Hisashi and Reed, Jason and Landry, Joseph W and Gewirtz, David A
Journal: Biochemical pharmacology (2019): 202-212
Authors: Saleh, Tareq and Tyutyunyk-Massey, Liliya and Murray, Graeme F and Alotaibi, Moureq R and Kawale, Ajinkya S and Elsayed, Zeinab and Henderson, Scott C and Yakovlev, Vasily and Elmore, Lynne W and Toor, Amir and Harada, Hisashi and Reed, Jason and Landry, Joseph W and Gewirtz, David A
Journal: Biochemical pharmacology (2019): 202-212
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