Xite™ Red beta-D-galactopyranoside
Xite™ Red beta-D-galactopyranoside provides a simple and sensitive tool to detect beta-galactosidase (β-gal) activity. Compared to the existing red beta-galactosidase substrates (e.g., the commonly used resorufin beta-D-galactopyranoside), it has much better cell permeability. Xite™ Red beta-D-galactopyranoside provides a simple and sensitive tool to detect beta-galactosidase activity. Xite™ Red 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. Xite™ Red beta-D-galactopyranoside enters readily cells where it gets cleaved by β-gal, producing Xite™ Red, a strongly fluorescent product. The strongly fluorescent Xite™ Red is well retained in cells, making it easy to be detected with a flow cytometer and fluorescence microscope. In addition, Xite™ Red beta-D-galactopyranoside is fixable. The red fluorescence generated by Xite™ Red beta-D-galactopyranoside can be readily combined with other color fluorescent probes such as DAPI or GFP for multicolor fluorescence analysis.
Example protocol
AT A GLANCE
Protocol summary
- Treat samples as desired.
- Prepare and add Xite™ Red 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 575/26 nm filter (PE channel) or using fluorescence microscopy with Cy3/TRITC 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™ Red beta-D-galactopyranoside stock solution
Add appropriate amount of DMSO into Xite™ Red beta-D-galactopyranoside to make 2-5 mM Xite™ Red beta-D-galactopyranoside stock solution. Note: Store the unused Xite™ Red beta-D-galactopyranoside stock solution at -20 °C in single use aliquots.PREPARATION OF WORKING SOLUTION
Xite™ Red beta-D-galactopyranoside working solution
Prepare 1-20 µM of Xite™ Red beta-D-galactopyranoside working solution in buffer of your choice. Note: Xite™ Red beta-D-galactopyranoside working solution should be used promptly. Note: The concentration of the Xite™ Red 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 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™ Red 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 575/26 nm filter (PE channel) or fluorescence microscope with Cy3/TRITC filter set.
References
View all 50 references: Citation Explorer
Acridinium Benzoates for Ratiometric Fluorescence Imaging.
Authors: Wen, Min and Wang, Xijing and Wang, Ting and Sun, Yan and Fan, Mengting and Li, Min and Zhu, Junru and Zhang, Dazhi and Cui, Xiaoyan and Shan, Yongkui
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2020): 3247-3251
Authors: Wen, Min and Wang, Xijing and Wang, Ting and Sun, Yan and Fan, Mengting and Li, Min and Zhu, Junru and Zhang, Dazhi and Cui, Xiaoyan and Shan, Yongkui
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2020): 3247-3251
Fluorescence Signal Amplification by Using β-Galactosidase for Flow Cytometry; Advantages of an Endogenous Activity-Free Enzyme.
Authors: Nobori, Takanobu and Kawamura, Masumi and Yoshida, Ryosuke and Joichi, Taisei and Kamino, Kenta and Kishimura, Akihiro and Baba, Eishi and Mori, Takeshi and Katayama, Yoshiki
Journal: Analytical chemistry (2020): 3069-3076
Authors: Nobori, Takanobu and Kawamura, Masumi and Yoshida, Ryosuke and Joichi, Taisei and Kamino, Kenta and Kishimura, Akihiro and Baba, Eishi and Mori, Takeshi and Katayama, Yoshiki
Journal: Analytical chemistry (2020): 3069-3076
Amphiphilic triphenylamine-benzothiadiazole dyes: preparation, fluorescence and aggregation behavior, and enzyme fluorescence detection.
Authors: Ishi-I, Tsutomu and Kawai, Kazuki and Shirai, Yuya and Kitahara, Ikumi and Hagiwara, Yoshinori
Journal: Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society (2019): 1447-1460
Authors: Ishi-I, Tsutomu and Kawai, Kazuki and Shirai, Yuya and Kitahara, Ikumi and Hagiwara, Yoshinori
Journal: Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society (2019): 1447-1460
Macrotheranostic Probe with Disease-Activated Near-Infrared Fluorescence, Photoacoustic, and Photothermal Signals for Imaging-Guided Therapy.
Authors: Zhen, Xu and Zhang, Jianjian and Huang, Jiaguo and Xie, Chen and Miao, Qingqing and Pu, Kanyi
Journal: Angewandte Chemie (International ed. in English) (2018): 7804-7808
Authors: Zhen, Xu and Zhang, Jianjian and Huang, Jiaguo and Xie, Chen and Miao, Qingqing and Pu, Kanyi
Journal: Angewandte Chemie (International ed. in English) (2018): 7804-7808
Impact of plasma protein binding on cargo release by thermosensitive liposomes probed by fluorescence correlation spectroscopy.
Authors: Mittag, Judith J and Kneidl, Barbara and Preiβ, Tobias and Hossann, Martin and Winter, Gerhard and Wuttke, Stefan and Engelke, Hanna and Rädler, Joachim O
Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenst (2017): 215-223
Authors: Mittag, Judith J and Kneidl, Barbara and Preiβ, Tobias and Hossann, Martin and Winter, Gerhard and Wuttke, Stefan and Engelke, Hanna and Rädler, Joachim O
Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenst (2017): 215-223
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