logo
AAT Bioquest

Cell Meter™ Cellular Senescence Activity Assay Kit *Green Fluorescence*

Cellular Senescence is an irreversible growth arrest triggered in order to prevent growth in DNA damaged cells. Senescence-associated beta-galactosidase (SA-beta-gal) is highly overexpressed in senescent cells and it has been widely used as a senescence marker. X-gal staining, a colorimetric method is widely available and used to detect SA-beta-gal in senescent cells. The color method has some limitations such as requirement of fixation of samples due to the low cell permeability of X-gal, longer staining time and low sensitivity. Cell Meter™ Cellular Senescence Activity Assay Kit uses Xite™ beta-D-galactopyranoside, a fluorogenic beta-Gal substrate that readily enters into live cells, and gets cleaved by SA-β-gal inside cells, generating strong green fluorescence. Unlike cell-impermeable X-Gal substrate, it has excellent cell permeability. Cell Meter™ Cellular Senescence Activity Assay Kit enables users to detect the senescence with higher sensitivity with robust performance. The Xite product is well retained inside the cells, producing a stable signal for fluorescence imaging and flow cytometry analysis.

Example protocol

AT A GLANCE

Protocol Summary
  1. Treat samples as desired
  2. Prepare and add Xite™ beta-D-galactopyranoside working solution to samples
  3. Incubate samples at 37 °C for 15 to 45 minutes
  4. Monitor the fluorescence intensity using flow cytometer with 530/30 nm filter (FITC channel)
Important Note

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

Xite™ beta-D-galactopyranoside stock solution (100X)

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

Xite™ beta-D-galactopyranoside working solution (1X)

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

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

Spectrum

Product family

Citations

View all 47 citations: Citation Explorer
TRAIP resolves DNA replication-transcription conflicts during the S-phase of unperturbed cells
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,
Journal: Nature Communications (2023): 5071
Merkel Cell Polyomavirus Large T Antigen Induces Cellular Senescence for Host Growth Arrest and Viral Genome Persistence through Its Unique Domain
Authors: Pham, Alexander M and Ortiz, Luz E and Lukacher, Aron E and Kwun, Hyun Jin
Journal: Cells (2023): 380
Topological DNA damage, telomere attrition and T cell senescence during chronic viral infections
Authors: Ji, Y., Dang, X., Nguyen, L. N. T., Nguyen, L. N., Zhao, J., Cao, D., Khanal, S., Schank, M., Wu, X. Y., Morrison, Z. D., Zou, Y., El Gazzar, M., Ning, S., Wang, L., Moorman, J. P., Yao, Z. Q.
Journal: Immun Ageing (2019): 12
Cell senescence, apoptosis and DNA damage cooperate in the remodeling processes accounting for heart morphogenesis
Authors: Lorda-Diez, C. I., Solis-Mancilla, M. E., Sanchez-Fern and ez, C., Garcia-Porrero, J. A., Hurle, J. M., Montero, J. A.
Journal: J Anat (2019): 815-829
Dynamic transcriptome profiling in DNA damage-induced cellular senescence and transient cell-cycle arrest
Authors: Zhao, Z., Dong, Q., Liu, X., Wei, L., Liu, L., Li, Y., Wang, X.
Journal: Genomics (2019): ersion="1.0" encoding="UTF-8" ?>23005.enlEndN
Page updated on December 13, 2024

Ordering information

Price
Unit size
Catalog Number23005
Quantity
Add to cart

Additional ordering information

Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
Technical SupportContact us
Purchase orderSend to sales@aatbio.com
ShippingStandard overnight for United States, inquire for international
Request quotation

Spectral properties

Absorbance (nm)

487

Correction Factor (260 nm)

0.32

Correction Factor (280 nm)

0.35

Extinction coefficient (cm -1 M -1)

800001

Excitation (nm)

498

Emission (nm)

517

Quantum yield

0.79001, 0.952

Storage, safety and handling

Intended useResearch Use Only (RUO)

Platform

Flow cytometer

Excitation488 nm laser
Emission530, 30 nm filter
Instrument specification(s)FITC channel

Fluorescence microscope

ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall, clear bottom

Components

Cellular senescence was measured with Cell Meter&trade; 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&trade; beta-D-galactopyranoside for 30 mins at 37<sup>o</sup>C. The signal was acquired using FITC channel in ACEA NovoCyte flow cytometer.
Cellular senescence was measured with Cell Meter&trade; 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&trade; beta-D-galactopyranoside for 30 mins at 37<sup>o</sup>C. The signal was acquired using FITC channel in ACEA NovoCyte flow cytometer.
Cellular senescence was measured with Cell Meter&trade; 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&trade; beta-D-galactopyranoside for 30 mins at 37<sup>o</sup>C. The signal was acquired using FITC channel in ACEA NovoCyte flow cytometer.
Cellular senescence was measured with Cell Meter&trade; 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&trade; beta-D-galactopyranoside for 30 mins at 37C. The signal was acquired using FITC filter set.
9L-LacZ cells&nbsp; were stained with DMSO or FDG or Xite&trade; 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.