The clickable mussel-derived peptides were prepared using standard Fmoc solid-phase synthesis47 (link). Quartz wafers (10 mm or 15 mm in diameter) with 80–100 nm TiO2 layer were prepared by the key Laboratory of Advanced Technologies of Materials, Southwest Jiaotong University (Chengdu, China). For in vivo experiments, pure Ti screws (2 mm × 10 mm) were purchased from Tianjin Zhengtian Medical Device Company (Tianjin, China). Cell counting kit-8 (CCK-8) kit, a Live/Dead cell staining kit and FITC-labeled Phalloidin staining were purchased from Yeasen Biotechnology Co. (Shanghai, China). Alkaline phosphatase (ALP) kit, ARS kit and Triton X-100 were obtained from Beyotime Biotechnology Co. (Jiangsu, China). The other chemical reagents or antibodies unless mentioned elsewhere were almost purchased from Sigma, Abcam or Invitrogen.
Live dead cell staining kit
Manufactured by Yeasen
Sourced in China
The Live/Dead cell staining kit is a laboratory tool designed to assess the viability of cells. It utilizes fluorescent dyes to distinguish between live and dead cells in a sample, providing a simple and effective way to evaluate cell health and population.
Automatically generated - may contain errors
Lab products found in correlation
Ars kit, by Beyotime (1 mentions)
Triton x 100, by Beyotime (1 mentions)
Alkaline phosphatase alp kit, by Beyotime (1 mentions)
Cell counting kit 8 cck 8 kit, by Yeasen (1 mentions)
Annexin 5 pi staining kit, by Beyotime (1 mentions)
Cck 8, by Apexbio (1 mentions)
Apoptosis analysis kit, by Yeasen (1 mentions)
Cell cycle analysis kit, by Yeasen (1 mentions)
4 protocols using live dead cell staining kit
1
Mussel-Derived Peptides for Bone Implant Optimization
2
Biocompatibility of GO/PCL Scaffolds
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To simulate potential cellular toxicity of the GO/PCL scaffold to in situ neural cells, primary SCs were utilized for biocompatibility testing instead of cell lines. Primary SCs were obtained by dissecting and digesting the sciatic nerve and brachial plexus nerve from neonatal rats (postnatal day 1) and undergoing passaging culture. GO/PCL scaffolds with varying GO concentrations (0.1%, 0.3%, 0.5%, 1.0%) were cut into 6 mm diameter discs using a puncher, sterilized with 75% ethanol for 2 hours, and then rinsed with 0.01 M PBS under ultraviolet light until the ethanol was completely removed (approximately 24 hours, 6 times). These scaffolds were placed onto 24-well plates and seeded with SCs at a density of 2 × 104 cells/mL (1 mL medium) in each well. The Live/Dead cell staining kit (Yeasen, China) was used for cell viability analysis according to the standard protocol. Calcein-AM and propidium iodide (PI) were prepared separately as 30 μM PBS solutions and added to cover the cells/scaffold completely. Following the removal of the working solution, the samples were washed with PBS three times and observed under an inverted fluorescence microscope.
3
Cell Viability and Apoptosis Assays
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Cell viability was assessed by a cell counting kit 8 (CCK-8; ApexBio, Houston, USA) assay according to the manufacturer’s protocol. The inhibition rate was calculated as follows: inhibition rate (x) = (ODcontrol - ODx)/ODcontrol. A Live/Dead cell staining kit (Yeasen) consisting of Calcein-AM (green fluorescence) and propidium iodide (PI, red fluorescence) and Annexin V/PI staining kit (Beyotime, Shanghai, China) was used according to the instructions to assess cell apoptosis.
4
PLGA Nanoparticle Synthesis and Characterization
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Poly(D,L-lactide-co-glycolide) (PLGA), polyvinyl alcohol (PVA), acetone, dichloromethane (DCM), and perfluoro-15-crown-5 were purchased from Sigma-Aldrich. Q1 (methyl(E)-3-[2-(3,4dihydroxyphenyl)-7-hydroxy-3-methoxycarbonyl-2,3-dihydro-1benzo furan-5yl] prop-2-enoate) was synthesized according to a previous report. 40 The cell counting kit (CCK-8) was purchased from MCE (USA), while the apoptosis analysis kit, cell cycle analysis kit and live/dead cell staining kit were purchased from Yeason (China).
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