Resveratrol (Trans-3, 4 ', 5-trihydroxystilnene), nucleosides (NS), and N-acetylcysteine (NAC) were purchased from Sigma-Aldrich (St. Louis, MO, United Stated). Phorbol-12-Myristate-13-Acetate (TPA) was purchased from Cell Signaling (Danvers, MA, United States). Dulbecco’s modified Eagle’s medium (DMEM) and other culture media were obtained from Invitrogen (Carlsbad, CA, United States). A senescence associated β-galactosidase (SA-β-gal) staining kit was purchased from Cell Signaling (Danvers, MA, United States). The antibodies used for the Western blot including p53, p21, Rad9, Slug, E-cadherin, and vimentin were purchased from Genetex (San Antonio, TX, United States). Antibodies such as γ-catenin, N-cadherin, and elongation factor 1 alpha (EF1A) were purchased from Abcam (Cambridge, UK). The antibodies including β-Actin, α-tubulin, and GAPDH were purchased from Proteintech Group Inc (USA). The HRP-conjugated goat anti-mouse and anti-rabbit secondary antibodies were purchased from Abcam (Cambridge, UK).
Senescence associated β galactosidase sa β gal staining kit
Manufactured by Cell Signaling Technology
Sourced in United States
The Senescence-associated β-galactosidase (SA-β-gal) staining kit is a laboratory tool used to detect and quantify senescent cells. It provides a simple, histochemical staining method to identify cells that express the senescence-associated β-galactosidase enzyme.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Resveratrol, by Merck Group (2 mentions)
Phorbol 12 myristate 13 acetate, by Cell Signaling Technology (1 mentions)
N acetylcysteine nac, by Merck Group (1 mentions)
N cadherin, by Abcam (1 mentions)
β actin, by Proteintech (1 mentions)
Gapdh, by Proteintech (1 mentions)
Vimentin, by GeneTex (1 mentions)
E cadherin, by GeneTex (1 mentions)
α tubulin, by Proteintech (1 mentions)
Glutathione (gsh), by Nanjing Jiancheng (1 mentions)
Mda detection kit, by Nanjing Jiancheng (1 mentions)
D galactose d gal, by Sangon (1 mentions)
Gsh px, by Nanjing Jiancheng (1 mentions)
Anti gpx1, by Abcam (1 mentions)
Horseradish peroxidase conjugated goat anti mouse secondary antibody, by Jackson ImmunoResearch (1 mentions)
2 7 dichlorodihydrofluorescein diacetate dcfh2 da, by Merck Group (1 mentions)
Anti col1, by Abcam (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
Fitc phalloidin, by Merck Group (1 mentions)
7 protocols using senescence associated β galactosidase sa β gal staining kit
1
Senescence Modulation by Bioactive Compounds
2
Aging Biomarker Quantification Protocol
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Rg1 (purity >95%) was purchased from Hongjiu Biotech Co., Ltd., (Tonghua, China). D-galactose (D-gal; purity >99%) was obtained from Sangon Biotech Co., Ltd. (Shanghai, China). The senescence-associated β-galactosidase (SA-β-gal) Staining kit was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), whereas the SOD, GSH-Px, GSH and MDA detection kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The AGEs (cat. no. abx512406) and 8-OH-dG (cat. no. SKT-120-480) kits were purchased from Shanghai Yuanye Biological Technology Co., Ltd. (Shanghai, China).
3
UVB-induced Skin Aging: Mechanisms and Mitigation
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Dulbecco's modified Eagle's medium (DMEM), penicillin, and streptomycin were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Fetal bovine serum (FBS) was obtained from GE Healthcare Life Sciences (Logan, UT, USA). UVB light (Philips 311 nm, TL 20W/01) was from Philips Lighting Holding B.V. (Eindhoven, The Netherlands). Cell Counting Kit-8 (CCK-8) was from Beyotime Institute of Biotechnology. RNase A, propidium iodide, FITC-phalloidin, and 2′,7′-dichlorodihydrofluorescein diacetate (DCFH2-DA) were from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Senescence-associated β-galactosidase (SA-β-gal) staining kit (#9860) was from Cell Signaling Technology Inc. (Danvers, MA, USA); terminal deoxynucleotidyl transferase- (TdT-) mediated dUTP-biotin nick end labeling (TUNEL) staining kit was from Roche Molecular Biochemicals (Mannheim, Germany). Anti-CD31 antibody was from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Horseradish peroxidase-conjugated goat anti-mouse antibody was from Dako (Glostrup, Denmark). Anti-GPX-1 (#ab108427), anti-catalase (#ab16731), anti-SOD-2 (#ab13533), anti-SOD-1 (#ab13498), anti-COL-1 (#ab6308), and β-actin (#ab6276) were from Abcam (Cambridge, UK); horseradish peroxidase-conjugated goat anti-mouse secondary antibody (#115-035-062) was from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA).
4
Senescence Evaluation of Adipose-Derived Mesenchymal Stem Cells
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Cellular senescence of ADMSCs was determined at the seventh passage of ADMSCs which had been cultured in medium containing different supplements including FBS, HPL, Hplasma, and HPL+Hplasma. Cells were seeded in a 35 mm dish at a density of 2.5×104 cells per dish in 2 ml of each medium and allowed to grow for three days. Cellular senescence was detected using a senescence-associated β-galactosidase (SA-β-gal) staining kit (Cell Signaling Technology, Inc., Danvers, Massachusetts) according to the manufacturer’s instructions. Briefly, cells were washed twice with PBS, fixed with 1x fixative solution for 15 minutes, and stained with the freshly prepared β-gal staining solution at a pH of 5.9 to 6.1. Dishes were sealed with parafilm to prevent evaporation and incubated at 37°C overnight in the absence of carbon dioxide. The presence of a blue colour, i.e. senescent cells, was observed under an inverted microscope. Ten fields of 10× magnification per dish were taken, and senescent cells were manually counted in each field. Data were presented as a percentage of senescent cells over the total number of cells.
5
Senescence Detection via SA-β-Gal Staining
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Senescence was detected with a senescence-associated-β-galactosidase (SA-β-Gal) staining kit (Cell Signaling Technology, Danvers, MA) following the manufacturer’s protocol as described in our previous study [14 (link)]. Images were obtained using an electron microscope (Olympus, Tokyo, Japan). The proportion of cells positive for SA-β-Gal staining is shown as the percentage of the total number of cells in each dish.
6
Cell Culture and Reagent Preparation
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Human lung adenocarcinoma cell line A549, human liver hepatocellular carcinoma cell line HepG2, murine macrophage cell line RAW 264.7, and primary human dermal fibroblast (HDF) cell line (Cell Applications, San Diego, CA, USA) were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Welgene, Gyeongsangbuk-do, Korea) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco, Waltham, MA., USA) and 1% penicillin/streptomycin (Corning Inc., Corning, NY., USA) at 37°C in a humid 5% CO2 atmosphere. Resveratrol was purchased from Sigma-Aldrich (St Louis, MO., USA), and the senescence-associated-β-galactosidase (SA-β-gal) staining kit was purchased from Cell Signaling Technology (Beverly, MA., USA).
7
Senescent Cardiomyocyte Identification
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Senescent cardiomyocytes were assessed using a senescence-associated β-galactosidase (SA-β-gal) staining kit (Cell Signaling Technology, USA). The cells were washed with phosphate-buffered saline three times. They were fixed for 15 min in a fixative solution at room temperature and then incubated overnight with a staining solution mix at 37°C. Senescent cells (SA-β-gal-positive) were identified under a microscope as blue-stained cells.
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