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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.
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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