“Senescent Cell Detection – Selection Guide”
Various Senescent Cell from Various Indicators
Product |
Cellular Senescence Detection Kit - SPiDER-ßGal |
Cellular Senescence Plate Assay Kit - SPiDER-ßGal |
DNA Damage Detection Kit – γH2AX * Coming Soon |
Nucleolus Bright Green / Red |
Detection |
Fluorescence |
Fluorescence |
Fluorescence |
Fluorescence |
Wavelength (Ex/Em) |
500 - 540 nm / 530 - 570 nm |
535 nm / 580 nm |
Green: 494 nm / 518 nm
Red: 550 nm / 566 nm
Deep Red: 646 nm / 668 nm
|
Green: 513 nm / 538 nm
Red: 537 nm / 605 nm
|
Indicator |
SA-ß-gal activity |
SA-ß-gal activity |
Changes in DNA damage |
Changes in the Nucleolus |
Detection method |
Imaging
Substrate:SPiDER-ßGal
|
Plate assay
Substrate:SPiDER-ßGal
|
Imaging Detection of γH2AX by secondary antibody method
|
Imaging Detection of the Nucleolus by RNA staining reagent
|
Instrument |
Fluorescence microscopy Flow cytometry
|
Plate reader |
Fluorescence microscopy |
Fluorescence microscopy |
Sample |
Live cells Fixed cells |
Live cells
(lysis of live cells)
|
Fixed cells |
Fixed cells |
data |
 |
 |
 |
 |
Item# |
SG03 |
SG05 |
Green: G265
Red: G266
Deep Red: G267
|
Green: N511
Red: N512 |
Feature of Cellular Senescence
Cellular senescence is considered crucial to many different research areas, especially since the recent discovery of the senescence-associated secretory phenotype (SASP). SASP is a known risk factor for malignant transformation and exploration into stem cell research has found a link between SASP and the aging phenomenon. The features of cellular senescence were summarized based on modern publications that have a high number of citations.
Evaluate senescent cells from various makers
In WI-38 cells at different passages SA-ß-Gal activity, mitochondrial membrane potential, and cellular metabolism (Glucose and Lactate) were evaluated using each specific kit. In senescent cells, SA-ß-Gal activity was enhanced and mitochondrial membrane potential was reduced. Consumption of Glucose and lactate level in the supernatant measured as an indicator of metabolism was increased.
Sample |
Senescence induction |
Senescence Marker |
Factor involved in senescence |
Citation |
IMR90 (human lung fibroblast cell) |
passage of aging |
SA-ßGal
p16, p21
enlarged nucleoli
|
SETD8 expression ↓
H4K20me1 ↓
mitochondrial oxidative phosphorylation ↑
ribosome biogenesis ↑
|
① |
SETD8 (methyltransferase) low expression |
mitochondrial oxidative phosphorylation ↑
ribosome biogenesis ↑
|
Aged satellite cells |
- |
SA-ßGal
p16
|
Autophagy activity ↓
ROS ↑
Mitochondria membrane potential ↓
|
② |
Atg7-deficient satellite cells |
Inhibition of autophagy |
SA-ßGal
P15, p16, p21
γ-H2AX
|
ROS ↑
Mitochondria membrane potential ↓
|
Fibroblasts from patients with type 2 diabetes |
- |
SA-ßGal
p21, p53
γ-H2AX
|
NADPH / NADP ↓
(resistance to oxidative stress ↓)
NADPH oxidase ↑
(ROS ↑)
|
③ |
IMR90 |
Ethidium Bromide (depletion of mtDNA)
+ pyruvate withdrawal
|
SA-ßGal |
NAD+ / NADH ↓ |
④ |
MDA-MB-231
(human breast cancer cells)
|
Radiation-induced senescence +
low expression of securin
|
SA-ßGal |
Lactate ↑
LDH activity ↑
(glycolysis ↑)
|
⑤ |
MEF
(mouse embryonic fibroblasts)
|
overexpression of cancer gene
passage of aging
overexpression of transcription factor(E2F1)
|
SA-ßGal
p16, p21
enlarged nucleoli
|
Ribosome RNA ↑
p53 ↑
|
⑥ |
Mouse tail-tip fibroblasts |
2 month-old WT,
22 month-old WT,
p16 KO(22 month-old)
|
SA-ßGal
p14, p16
|
NAD+ ↓
SIRT3 ↓
|
⑦ |
Publications
①
H. Tanaka, S. Takebayashi, A. Sakamoto, N. Saitoh, S. Hino and M. Nakao, “The SETD8/PR-Set7 Methyltransferase Functions as a Barrier to Prevent Senescence-Associated Metabolic Remodeling.”, Cell Reports, 2017, 18(9), 2148.
②
L. Garcia-Prat, M. Martinez-Vicente and P. Munoz-Canoves, “Autophagy: a decisive process for stemness”, Oncotarget, 2016, 7(11), 12286.
③
M. Bitar, S. Abdel-Halim and F. Al-Mulla, “Caveolin-1/PTRF upregulation constitutes a mechanism for mediating p53-induced cellular senescence: implications for evidence-based therapy of delayed wound healing in diabetes”, Am J Physiol Endocrinol Metab., 2013, 305(8), E951.
④
C. Wiley, M. Velarde, P. Lecot, A. Gerencser, E. Verdin, J. Campisi, et. al., “Mitochondrial Dysfunction Induces Senescence with a Distinct Secretory Phenotype”, Cell Metab., 2016, 23(2), 303.
⑤
E. Liao, Y. Hsu, Q. Chuah, Y. Lee, J. Hu, T. Huang, P-M Yang & S-J Chiu, “Radiation induces senescence and a bystander effect through metabolic alterations.”, Cell Death Dis., 2014, 5, e1255.
⑥
K. Nishimura, T. Kumazawa, T. Kuroda, A. Murayama, J. Yanagisawa and K. Kimura, “Perturbation of Ribosome Biogenesis Drives Cells into Senescence through 5S RNP-Mediated p53 Activation”, Cell Rep. 2015, 10(8), 1310.
⑦
M. J. Son, Y. Kwon, T. Son and Y. S. Cho, “Restoration of Mitochondrial NAD+ Levels Delays Stem Cell Senescence and Facilitates Reprogramming of Aged Somatic Cells”, Stem Cells. 2016, 34(12), 2840.