Cell Meter™ Cellular Senescence Activity Assay Kit *Green Fluorescence*
![Cellular senescence was measured with Cell Meter™ Cellular Senescence Activity Assay Kit using a NovoCyte Flow Cytometer (ACEA Biosciences). HL-60 cells were incubated with Camptothecin for 6 hours to induce senescence and stained with Xite™ beta-D-galactopyranoside for 30 mins at 37<sup>o</sup>C. The signal was acquired using FITC channel in ACEA NovoCyte flow cytometer.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-meter-cellular-senescence-activity-assay-kit%2Ffigure-for-cell-meter-cellular-senescence-activity-assay-kit_j0oKR.jpg&w=640&q=75)
![Cellular senescence was measured with Cell Meter™ Cellular Senescence Activity Assay Kit using a NovoCyte Flow Cytometer (ACEA Biosciences). HL-60 cells were incubated with Camptothecin for 6 hours to induce senescence and stained with Xite™ beta-D-galactopyranoside for 30 mins at 37<sup>o</sup>C. The signal was acquired using FITC channel in ACEA NovoCyte flow cytometer.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-meter-cellular-senescence-activity-assay-kit%2Ffigure-for-cell-meter-cellular-senescence-activity-assay-kit_j0oKR.jpg&w=640&q=75)
![Cellular senescence was measured with Cell Meter™ Cellular Senescence Activity Assay Kit using a NovoCyte Flow Cytometer (ACEA Biosciences). HL-60 cells were incubated with Camptothecin for 6 hours to induce senescence and stained with Xite™ beta-D-galactopyranoside for 30 mins at 37<sup>o</sup>C. The signal was acquired using FITC channel in ACEA NovoCyte flow cytometer.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-meter-cellular-senescence-activity-assay-kit%2Ffigure-for-cell-meter-cellular-senescence-activity-assay-kit_j0oKR.jpg&w=128&q=25)
![Cellular senescence was measured with Cell Meter™ Cellular Senescence Activity Assay Kit using a fluorescence microscope. HeLa cells were incubated with Camptothecin for 6 hours to induce senescence and stained with Xite™ beta-D-galactopyranoside for 30 mins at 37C. The signal was acquired using FITC filter set.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-meter-cellular-senescence-activity-assay-kit%2Ffigure-for-cell-meter-cellular-senescence-activity-assay-kit_TOKkH.jpg&w=128&q=25)
![9L-LacZ cells were stained with DMSO or FDG or Xite™ beta-D-galactopyranoside for 30 minutes at 37C incubator, an then the cells were washed twice and scrapped using a scrapper with 1 mL HH buffer. Cells were collected and analyzed with flow cytometer using FITC channel.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-meter-cellular-senescence-activity-assay-kit%2Ffigure-for-cell-meter-cellular-senescence-activity-assay-kit_n81XB.jpg&w=128&q=25)
AT A GLANCE
- Treat samples as desired
- Prepare and add Xite™ 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)
Thaw each kit component at room temperature before starting the experiment.
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
Add 100 uL DMSO (Component C) into Xite™ beta-D-galactopyranoside (Component A) and mix well. Note: Store the unused Xite™ beta-D-galactopyranoside stock solution at -20 °C in single use aliquots.
PREPARATION OF WORKING SOLUTION
Dilute 10 uL of Xite™ beta-D-galactopyranoside stock solution (100X) with 1 mL of Assay Buffer to make Xite™ beta-D-galactopyranoside working solution (1X). Note: Xite™ beta-D-galactopyranoside working solution should be used promptly.
SAMPLE EXPERIMENTAL PROTOCOL
Treat your samples as desired.
Note: For 96 well plate format, grow cells in 100 µL of cell culture medium and treat as desired. Volumes can be adjusted based on the plate size.
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 100 µL Xite™ 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 carefully.
Remove the working solution and wash cells with buffer of your choice.
Note: If performing flow cytometry for attached cells, cells can be trypsined at this step and collected in a tube.
- Resuspend the cells in the Assay Buffer (Component B) and monitor the fluorescence intensity with flow cytometer using 530/30 nm filter (FITC channel) or fluorescence microscope with FITC filter set.
Name | Excitation (nm) | Emission (nm) |
Cell Meter™ Cellular Senescence Activity Assay Kit *Red Fluorescence* | 544 | 567 |
Authors: Scaramuzza, Shaun and Jones, Rebecca M and Sadurni, Martina Muste and Reynolds-Winczura, Alicja and Poovathumkadavil, Divyasree and Farrell, Abigail and Natsume, Toyoaki and Rojas, Patricia and Cuesta, Cyntia Fernandez and Kanemaki, Masato T and others,
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