After 24 h of cell treatment, the walled cells were trypsinized again, and the terminated digested cells were subjected to induction of apoptosis according to the instructions of Apoptosis Detection Kit (E‐CK‐A211, Elabscience). The cells were collected by centrifugation at 1000 rpm for 5 min, the supernatant was discarded, and the cells were washed twice with pre‐chilled PBS to adjust the number of cells per tube to 0.2–1.0 × 106. Add 400 µL 1× binding buffer to resuspend the cells. Add 5 µL FITC‐Annexin V to each tube and react at room temperature for 15 min; then add 10 µL PI and mix lightly for 5 min at 4°C. After the reaction, complete the assay in BD Accuri C6 device (BD) within 1 h. The appropriate channels were selected, and additional blank tubes (without fluorescent reagent) and PI single‐stained tubes (with PI reagent only) were set up for voltage adjustment of the flow instrument and compensation of fluorescent channels, and the results were analysed by FlowJo 7.6 software.
Apoptosis detection kit
Manufactured by Elabscience
Sourced in China
The Apoptosis Detection Kit is a laboratory tool designed to detect and analyze apoptosis, a form of programmed cell death. The kit provides the necessary reagents and protocols to identify and quantify apoptotic cells through specific cellular and molecular markers. The core function of this product is to facilitate the study of apoptosis, a fundamental biological process, in various experimental and research settings.
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Lab products found in correlation
Accuri c6 device, by BD (1 mentions)
Facsverse calibur ariaiisorp, by BD (1 mentions)
Intracellular atp assay kit, by Beyotime (1 mentions)
Mitosox red, by Thermo Fisher Scientific (1 mentions)
Mannitol, by Merck Group (1 mentions)
Horseradish peroxidase hrp substrate, by Merck Group (1 mentions)
M pertm mammalian protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
Thiazolyl blue tetrazolium bromide mtt, by Bio Basic (1 mentions)
Dulbecco s modified eagle s medium dmem, by Nacalai Tesque (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Novaseq 6000 sp reagent kit v1, by Illumina (1 mentions)
5 protocols using apoptosis detection kit
1
Quantifying Apoptosis in Treated Cells
2
Apoptosis Quantification in PTC Cells
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At 48 h posttransfection, the PTC cells were harvested, and cell apoptosis was determined by using the apoptosis detection kit purchased from Elabscience Biotechnology (Shanghai, China) according to the provided experimental procedures. In brief, the cells were diluted by using the DMEM at a density of 2 × 106 mL−1, 0.5 mL of cell suspensions was centrifuged at 1000 g for 5 min, and the cells were collected. After that, the cells were suspended in the same volume (0.5 mL) of precooled 1× binding buffer, followed by 15‐min incubation under dark conditions with 5 μL Annexin V–FITC and 10 μL propidium iodide (PI). Finally, a flow cytometer (FACSVerse/Calibur/AriaIISORP; BD Biosciences, San Jose, CA, USA) was used to measure the apoptosis ratio of cells.
3
Investigating Mitochondrial Function
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Z-LIG (purity ≥ 98%, product number: PS010302, CAS number: 81944-09-4) was purchased from Push Bio-technology (Chengdu, China). The Apoptosis Detection Kit was obtained from Elabscience (Wuhan, China). The Intracellular ATP Assay Kit and JC-1 were purchased from Beyotime (Shanghai, China). MitoSOX™ Red was obtained from Invitrogen (Carlsbad, CA, USA). Details of the antibodies used are listed in Additional file 1 : Table S1. The primer sequences are summarized in Additional file 2 (Tables S2) and Additional file 3 (Table S3). Unless otherwise specified, all other chemicals were purchased from Sigma-Aldrich (St, Louis, MO, USA).
4
Alginate-Based Nanoparticle Synthesis and Characterization
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Sodium alginate (300 – 400 mPa⋅s) was purchased from Chemiz (Malaysia). Gold (III) chloride trihydrate (HAuCl4·3H2O), 3,3′-anthracene-9,10-diyldipropanoic acid (ADPA), 1,3-diphenylisobenzofuran (DPBF), triton X-100 and mannitol were purchased from Sigma-Aldrich (Germany). Sodium borohydride (NaBH4), dimethyl sulfoxide (DMSO) and horseradish peroxidase (HRP) substrate were supplied by Merck (Germany). M-PERTM mammalian protein extraction reagent was purchased from Thermo Fischer Scientific (USA). Thiazolyl blue tetrazolium bromide (MTT) and ascorbic acid were obtained from BioBasic (Canada). Hexadecyltrimethylammonium bromide (CTAB) was purchased from Sangon Biotech (China). Silver nitrate (AgNO3) was acquired from Bendosen (Malaysia). Dulbecco’s modified Eagle’s medium (DMEM) was purchased from Nacalai Tesque (Japan). Fetal bovine serum (FBS) was purchased from GIBCO™ (USA). Apoptosis detection kit was supplied by Elabscience (USA). Unless stated otherwise, all reagents were prepared using Milli-Q ultrapure water.
5
Multi-Omics Analysis of Apoptosis
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BLM was purchased from Selleckchem (Houston, Texas, United States). Apoptosis detection kit was collected from Elabscience Biotechnology Co., Ltd. (Wuhan, China). RNA isolation kit was collected from Chengdu Foregene Biotechnology Co., Ltd. (Chengdu, China). Removal kit for genomic DNA and EvaGreen Express qPCR MasterMix-No Dye were obtained from ABM (Vancouver, Canada). Primer sequence synthesis was conducted by Sangon Biotech (Shanghai, China). DNA Microprep Kit and Zymoclean Gel Recovery Kit were obtained from Zymo Research BIOMICS (California, United States). Prep Kit of DNA Library for Illumina was obtained from BioLabs (United States). NovaSeq 6000 SP Reagent Kit V1.5 was obtained from Illumina (California, United States). In Situ Cell Death Detection Kit (TMR red fluorescence) was obtained from Roche Diagnostics (Germany). 3,3′-Diaminobenzidine (DAB) kit was collected from MXB biotechnologies (China). Cleaved caspase-3 and BAX antibodies were collected from CST (United States) and Boster (United States), respectively.
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