HOS and 143B cells were seeded in 24-well plates (2.5 × 104/well) and cultured overnight. Cis (20 μmol/L), UA (35 μmol/L), or Cis (20 μmol/L)+UA (35 μmol/L) was added to the wells for 24 h. Then, a calcein-AM/PI staining kit (Beyotime, Shanghai, China) was used to detect dead and live cells. PI was used to stain dead cells, and calcein-AM was used to stain live cells.
Calcein am pi staining kit
Manufactured by Beyotime
Sourced in China
The Calcein-AM/PI staining kit is a fluorescent dye-based solution used to detect cell viability and cytotoxicity in cell cultures. The kit contains Calcein-AM, a cell-permeant dye that is converted to a green fluorescent compound by live cells, and Propidium Iodide (PI), a red fluorescent dye that stains the nuclei of dead or membrane-compromised cells. This combination allows for the simultaneous identification of live and dead cells within a sample.
Automatically generated - may contain errors
Lab products found in correlation
Cell counting kit 8 cck 8 kit, by Beyotime (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Cisplatin, by Yuanye Bio-Technology (1 mentions)
Copper acetate, by Maclin Biochemical Technology (1 mentions)
17β estradiol e2, by Cayman Chemical (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Annexin 5 fitc pi, by Beyotime (1 mentions)
Hoechst 33342, by Beyotime (1 mentions)
Gsh assay kit, by Beyotime (1 mentions)
Nitric oxide assay kit, by Nanjing Jiancheng (1 mentions)
Lyso tracker red, by Beyotime (1 mentions)
H2o2 assay kit, by Solarbio (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Perfect real time rt reagent kit, by Takara Bio (1 mentions)
Phosphate buffer pbs, by Solarbio (1 mentions)
Alizarin red stain, by Cyagen (1 mentions)
Osteopontin opn, by Abcam (1 mentions)
Polyvinyl alcohol (pva), by Merck Group (1 mentions)
Eastep super total rna extraction kit, by Promega (1 mentions)
Paraformaldehyde (pfa), by Solarbio (1 mentions)
7 protocols using calcein am pi staining kit
1
Apoptosis Induction in Tumor Cells
2
Copper-based Nanoparticle Anticancer Protocol
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Copper acetate was obtained from Macklin. Gallic acid (GA) was purchased from Kermel. Surfactant Aerosol OT was obtained from Alfa Aesar. Cisplatin was purchased from Shanghaiyuanye Bio-Technology Co., Ltd. l -arginine (l -Arg), NADH, glutathione (GSH) and H2O2 Assay Kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. DSPE-PEG2000 was purchased from Top-Peptide Co., Ltd. BCA Kit, Lyso-Tracker Red, Hoechst 33342, DAPI, GSH Assay Kit, Calcein-AM/PI staining kit and Annexin V-FITC/PI were obtained from Beyotime Biotechnology. Charcoal-stripped fetal bovine serum (CS-FBS) was supplied by Hyclony. Nitric Oxide Assay Kit was obtained from Nanjing Jiancheng Bioengineering Institute. NO fluorescence probe (DAF-FM DA) was purchased from meilunbio. 4-hydroxyestradiol (4-OHE2) and 17β-estradiol (E2) were obtained from Cayman Chemical. PD98059 and Calpeptin were purchased from MCE. All other chemicals were provided by Sigma Aldrich (St. Louis, MO, USA) unless mentioned otherwise.
3
Rabbit BMSC Osteogenic Differentiation
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Sodium tetraborate (Na2B4O7.10H2O) was purchased from Beijing Chemical Works (Beijing, China). PVA (≈99% hydrolyzed, Mw ≈130,000) and TEOS were obtained from Sigma-Aldrich. The rabbit BMSCs were supplied by American Type Culture Collection (ATCC, MD, United States). Low-glucose Dulbecco’s modified Eagle’s medium (LG-DMEM), fetal bovine serum (FBS), and streptomycin–penicillin were obtained from Gibco® Life Technologies (CA, United States). The Cell Counting Kit-8 (CCK-8) kit and Calcein-AM/PI staining kit were supplied by Beyotime Biotechnology (Shanghai, China). The mediums for osteogenic differentiation of BMSCs and Alizarin Red stain were obtained from Cyagen Biosciences (CA, United States). Phosphate buffer (PBS) and 4% paraformaldehyde were provided by Solarbio (Beijing, China). The Eastep Super Total RNA Extraction Kit was purchased from Promega (Shanghai, China), and the Perfect Real-Time RT reagent kit was supplied by Takara Bio (Dalian, China). Hematoxylin–eosin stain was obtained from Thermo Fisher Scientific, MA, United States. Primary antibodies, runt-related transcription factor-2 (Runx-2), type I collagen (Col-1), osteocalcin (OCN), and osteopontin (OPN) were purchased from Abcam (Cambridge, United Kingdom), and secondary antibodies were obtained from Jackson ImmunoResearch Laboratories (West Grove, PA, United States).
4
Plumbagin's Cytotoxic Effects on Liver Cancer Cells
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Huh-7 and Hep-G2 cells were plated in 96-well plates (2 × 103/well) and treated with plumbagin at indicated concentrations for 12 h. Cell viability was measured using cck-8 kit (GlpBio, Montclair, CA, USA) following the manufacturer’s instructions, and the absorbance of each well was measured at 450 nm using a microplate reader (Thermo, Waltham, MA, USA). The half-maximal inhibitory concentration (IC50) was calculated.
Calcein-AM/PI staining kit (Beyotime, Shanghai, China) was used for cell viability/cytotoxicity detection. In brief, Huh-7 and Hep-G2 cells were plated on 24-well plates (3000 cells per well) and incubated overnight. Then, cells were treated by PLB at indicated concentrations for 12 h. After treatment, cells were washed by PBS and stained with Calcein-AM/PI detection buffer for 30 min. After staining, cells were washed twice with PBS buffer and observed under fluorescence microscope (Olympus, Tokyo, Japan).
Calcein-AM/PI staining kit (Beyotime, Shanghai, China) was used for cell viability/cytotoxicity detection. In brief, Huh-7 and Hep-G2 cells were plated on 24-well plates (3000 cells per well) and incubated overnight. Then, cells were treated by PLB at indicated concentrations for 12 h. After treatment, cells were washed by PBS and stained with Calcein-AM/PI detection buffer for 30 min. After staining, cells were washed twice with PBS buffer and observed under fluorescence microscope (Olympus, Tokyo, Japan).
5
Cell Viability Determination of SVF-derived Cells
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The SVF-gel and TE-SVF-gel was first liquefied by adjusting the temperature to 37°C in incubator. Following liquefaction, the SVF-gel and TE-SVF-gel was added to saline solution. The mixture was then centrifuged at 300 g for 5 min to collect the cells. The viability of the cells was assessed using a Calcein-AM/PI staining kit (Beyotime). In brief, cells were washed with PBS and then incubated with a staining solution containing Calcein-AM and propidium iodide (PI) in phosphate-buffered saline (PBS) for 30 min at 37°C in a humidified atmosphere of 5% CO2. Following incubation, cells were washed twice with PBS to remove excess dye. Live cells, characterized by their ability to convert non-fluorescent calcein-AM into green-fluorescent calcein, and dead cells, identified by their permeability to PI which intercalated into DNA and RNA to emit red fluorescence, were visualized under a confocal fluorescence microscope. The number of live and dead cells was counted in three random fields per well, and the percentage of viable cells was calculated as (number of live cells/total number of cells) × 100% (Liu et al., 2020 (link)).
6
Biomimetic 3D Scaffold for Bone Tissue Engineering
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GelMA, HAMA and lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP) were purchased from SunP Boyuan Biotech Co., Ltd (Beijing, China). NHAP was purchased from Sigma-Aldrich (MO, USA). Osteogenic induction medium and Alizarin Red Staining (ARS) kit were purchased from Cyagen Bioscienes (Guangzhou, China). Fetal bovine serum was purchased from Moregate Biotech Co., Ltd (Bulimba, Australia). Dulbecco's modified Eagle's medium (DMEM) and Trypsin were obtained from Gibco (Grand Island, USA). DMEM/F-12 supplemented with the REGM SingleQuot kit was bought from Lonza (USA). All the Cytokines were bought from Peprotech (Rocky Hill, USA). All the antibodies were bought from Proteintech Group Inc (Wuhan, China). Crystalline violet staining kit, DAPI and FITC were purchased from Solarbio Life Sciences (Beijing, China). Total protein assay kit, Cell Counting Kit-8(CCK8) kit, Alkaline Phosphatase (ALP) staining kit, DiI Fluorescent Staining Kit, DiR Fluorescent Staining Kit and Calcein-AM/PI staining kit was purchased from Beyotime Biotechnology (Shanghai, China). Crystalline violet reagent purchased from Solarbio Life sciences (Beijing, China). Growth factor-reduced Matrigel was purchased from BD Bioscience (Shanghai, China). SteadyPure Universal RNA Extraction Kit, Evo M-MLV RT Premix Kit and SYBR® Green Premix Pro Taq HS qPCR Kit were purchased from Accurate Biology (Changsha, China).
7
Apoptosis Induction in PANC-1 Cells
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PANC-1 cells were seeded in 6-well plates (2 × 105/well) and cultured for 24 h. DHA (40 µM) and DDP (40 µM) were added to the indicated culture plate wells for 24 h. Then, cells were harvested and subjected to Calcein-AM/PI staining kit (Beyotime, Shanghai, China) at 37 °C for 15 min. Nuclei were counterstained with DAPI (10 μg/mL) for 5 min. Fluorescence microscope (Nikon) was employed to take photos; PI stained for the dead cells, and Calcein AM stained for the live cells. Treated cells were also incubating with 5 μL Annexin V-FITC and 10 μL PI for 5 min at room temperature in the dark according to the manufacturer’s instructions (MultiSciences, Hangzhou, China). Single-cell suspensions were subjected to flow cytometry (ACEA NovoCyte, USA) and the percentage of Live/Dead cells was quantified.
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