The following materials were purchased from Thermo Fisher Scientific, Inc.: Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), penicillin, streptomycin, dimethyl sulfoxide (DMSO), TRIzol reagent and chemiluminescence reagents. The following materials were purchased from Beyotime Institute of Biotechnology: Senescence Cells Histochemical Staining kit (cat. no. C0602), Cell Counting Kit-8 (CCK-8; cat. no. C0038), RIPA lysis buffer, BCA protein assay kit and polyvinylidene difluoride (PVDF) membrane. PrimeScript RT Master mix kit and SYBR Green Master Mixture were from Takara Biotechnology Co., Ltd. Hnk (cat. no. HY-N0003) was obtained from MedChemExpress. Doxorubicin hydrochloride (cat. no. D1515) was from Sigma-Aldrich; Merck KGaA. Anti-p16INK4A (cat. no. ARG57377) was from Arigo Biolaboratories. Anti-p21 (cat. no. ab109199) was from Abcam. Anti-TXNIP (cat. no. sc-166234) was from Santa Cruz Biotechnology, Inc. The β-tubulin antibody (cat. no. 66240-1-Ig) was from ProteinTech Group, Inc. Secondary antibodies conjugated to horseradish peroxidase and enhanced chemiluminescence reagents were from AntGene Biotechnology Co., Ltd.
Senescence cells histochemical staining kit
Manufactured by Beyotime
Sourced in China
The Senescence Cells Histochemical Staining kit is a laboratory tool designed to detect and quantify senescent cells. It utilizes a histochemical staining method to identify the presence of senescence-associated beta-galactosidase (SA-β-gal) activity, a widely recognized marker of cellular senescence.
Automatically generated - may contain errors
Lab products found in correlation
Primescript rt master mix kit, by Takara Bio (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
β tubulin antibody, by Santa Cruz Biotechnology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Sybr green master mixture, by Takara Bio (1 mentions)
6 protocols using senescence cells histochemical staining kit
1
Senescence-associated Biomarker Expression
2
Senescence Cells Histochemical Staining
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EPCs were stained using a Senescence Cells Histochemical Staining kit (Beyotime Institute of Biotechnology, Shanghai, China, C0602) to assess senescence [24 (link)]. Briefly, the cells were washed with PBS three times and then incubated for 18 h in SA-β-gal staining solution at 37 °C with no CO2. Then, the samples were washed again with PBS three times and cover-slipped for direct imaging and counting under a microscope.
3
Senescence Assay for BMSCs and iPSC-MSCs
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In order to study the senescence of the BMSCs (Passage 4) and iPSC-MSCs (Passage 4), the Senescence Cells Histochemical Staining Kit (Beyotime Biotechnology, China) was used in accordance with the manufacturer’s instructions. Random images were photographed during observation under a microscope (Olympus IX71, Japan). The number of cells with intracellular blue deposits divided by the total number of cells was the ratio of senescent cells. Five independent experiments were performed to ensure reproducibility.
4
Detecting Cellular Senescence through SA-β-gal and Cell Cycle Inhibitors
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To detect senescence, two different features of senescence were investigated: i) Enhanced β-galactosidase activity (SA-β-gal) associated with increased lysosomal content; and ii) expression of the cell cycle inhibitor p16INK4A and p21 (35 (link)). Staining for SA-β-gal was performed with a Senescence Cells Histochemical Staining kit (Beyotime Institute of Biotechnology), according to the manufacturer's protocols. Briefly, cultured H9c2 cardiomyocytes were fixed in 4% formaldehyde for 15 min at room temperature and then washed three times in phosphate-buffered saline (PBS) at room temperature. The slides were immersed in freshly prepared SA-β-gal staining solution and incubated at 37°C without CO2 overnight. Stained sections were washed twice with PBS. Senescence was quantitated by visual inspection of blue/green stained cells with an inverted microscope (magnification ×200). In total, >6 observations were included in each group.
5
Senescence Cell Histochemical Staining
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EPCs were stained using a Senescence Cells Histochemical Staining kit (Beyotime Institute of Biotechnology), according to the manufacturer’s protocols, to assess senescence. Senescence was quantitated by visual inspection of blue/green stained cells with an inverted microscope (magnification, ×10). More than 4 mice were included in each group.
6
Senescence Cell Histochemical Staining
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The senescence Cells Histochemical Staining Kit (Beyotime C0602) was used to evaluate senescence following manufacturer's instructions. Firstly, cells or tissues were incubated with fixation buffer for 10 minutes at room temperature. And then, samples were washed and incubated with staining mixture (containing X‐gal) at 37°C for 24‐30 hours. Senescence degree was next detected by a fluorescence microscope (Leica Dmi8, Germany). Finally, the numbers of SA‐β‐gal‐positive cells were counted by ImageJ software in a blinded manner.
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