Glass-bottomed Petri dishes were used to seed rBMSCs and incubated overnight. The vehicles loaded with FAM-labeled miRNA molecules were added with a final concentration of 20 μg/mL. After allowing different incubation time periods, the medium was replaced with the prewarmed (37°C) Lyso-Tracker Red (Solarbio Life Sciences) probe-containing medium to label the lysosome. The loading solution was removed after an hour and the cells were washed twice with PBS. They were then fixed with 4% paraformaldehyde and washed twice with PBS; the cells were dyed with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime Biotechnology) to label the cell nuclei. Fluorescence images were captured using CLSM (Zeiss-LSM710; Carl Zeiss Meditec AG, Jena, Germany).
Lyso tracker red
Manufactured by Solarbio
Sourced in China
The Lyso-Tracker Red is a fluorescent dye that specifically labels acidic organelles, such as lysosomes, in live cells. It is a useful tool for studying lysosomal function and dynamics in a variety of cell types.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Solarbio (3 mentions)
Lsm 710, by Zeiss (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
2 7 dichlorofluorescin diacetate dcfda, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Anhydrous dimethyl sulfoxide dmso, by Merck Group (1 mentions)
Cerium 3 acetate hydrate, by Merck Group (1 mentions)
Catalase activity assay kit, by Solarbio (1 mentions)
Cd206 antibody, by BioLegend (1 mentions)
Cd68 antibody, by BioLegend (1 mentions)
Urea, by Sinopharm (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Celltracker green cmfda, by Yeasen (1 mentions)
N 3 dimethylaminopropyl n ethylcarbodiimide hydrochloride edc, by Merck Group (1 mentions)
N hydroxysuccinimide nhs, by Merck Group (1 mentions)
6 diamidino 2 phenylindole dapi, by Merck Group (1 mentions)
Resveratrol, by Merck Group (1 mentions)
Mitotracker red, by Solarbio (1 mentions)
Calcein am pi kit, by Solarbio (1 mentions)
16 protocols using lyso tracker red
1
Intracellular Trafficking of miRNA
2
Fluorescent Nanoparticle Synthesis and Characterization
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Tetraethyl orthosilicate (TEOS), Cetyltrimethylammonium bromide (CTAB) were obtained from Sinopharm Chemical Reagent Co. Urea, ferric acetylacetonate (Fe(acac)3), fluorescein isothiocyanate (FITC), Cerium (III) acetate hydrate, anhydrous dimethylsulfoxide (DMSO), 2′,7′-Dichlorofluorescin diacetate (DCF-DA) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich (USA). mPEG-NH2 (5kD) was purchased from Aladdin (China). Sodium salicylate (NaSal), triethanolamine (TEA), Cell Counting Kit-8 (CCK-8) were purchased from MedChemExpress (MCE, USA). Penicillin, streptomycin, fetal bovine serum (FBS), and Dulbecco’s Modified Eagle’s Medium (DMEM) were purchased from Hyclone (USA). 4′, 6-Diamidino-2-phenylindole (DAPI), Lyso-Tracker Red, total superoxide dismutase assay kit with WST-8, and catalase activity assay kit were purchased from Solarbio (China). CD68 antibody, Alexa Fluor 549 anti-Nos2 (iNOS) antibody, and CD206 antibody were purchased from Biolegend (USA). Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) ELISA kit were purchased from FineTest (China).
3
Phagocytosis of Apoptotic Thymocytes
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Strain-matched thymi were filtered through a 200-mesh screen to collect single cells, and the liberated thymocytes were stained with Cell Tracker Green CMFDA (40721ES60, Yeasen, Shanghai, China). Then, thymocyte apoptosis was induced by overnight culture in RPMI 1640 with the final 1 μmol/L of dexamethasone in (50–02–2, Sigma-Aldrich). BMDMs were seeded in a 24-well plate and incubated with Lyso-Tracker Red (L8010, Solarbio, Beijing, China) for 15 min. Macrophages were then overlaid with apoptotic thymocytes at a ratio of 10:1 for 1 h. After three times of washing with ice-cold PBS, colocalization between the red phagolysosome and green apoptotic thymocytes to represent phagocytosis using fluorescence microscopy.
4
Graphene Oxide-Based Theranostic Nanoplatform
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Graphene oxide powder was purchased from Nanjing XF NANO Materials Tech Co., Ltd (China). Mitotracker red, Lysotracker red, Calcein AM/PI kit, Annexin V-FITC/PI apoptosis staining kit were purchased from Solarbio (Beijing, China). IR-780 iodide (IR780), ferritin (FRT) from equine spleen, resveratrol (RSV, ≥99%), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimidehydrochloride (EDC), N-hydroxysuccinimide (NHS) and 6-diamidino-2-phenylindole (DAPI) were purchased from Sigma-Aldrich (USA). All other chemicals used in the study were from Aladdin (Shanghai, China).
5
Cellular Uptake and Localization of pCas12f-FITC Complexes
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MCF-7 and HEK293T cells were seeded on 24 mm2 glass coverslips in a 6-well plate and treated with TSPscp/pCas12f-FITC, TSP/pCas12f-FITC, PEI/pCas12f-FITC and Lipo/pCas12f-FITC complexes at a concentration of 500 ng pCas9 per well for 8 h. The cells were stained with LysoTracker Red (Solarbio) for 1 h and 4’,6-diamidino-2-phenylindole (DAPI, H3570, Invitrogen) for 3 min, washed with PBS three times, and finally imaged using a confocal laser scanning microscope (CLSM, A1, Nikon) to visualize the cellular uptake and localization of the pCas12f-FITC complexes.
6
SARS-CoV-2 Protein Interaction and Cellular Uptake
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BMDCs, RAW264.7 and Vero cells were seeded in six-well dish and treated with 10 μg of RBD-Trimer, COE-Trimer, S1-Trimer, RBD-Monomer, COE-Monomer or S1-Monomer for 6 h. Lyso-Tracker Red (Solarbio, China; 1:15000), FITC-Monoclonal Mouse Anti-His antibody (OKA, China; 1:1000), and 4′,6-diamidino-2-phenylindole (DAPI; Sigma, United States; 1:2000) were used to label lysosomes, antigens, and nuclei. An inverted fluorescence microscope was used to take fluorescence images (Olympus, Japan). CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) (Promega, United States) was performed to detect cell viability. Optical density (OD) was measured at 490 nm.
7
Multimodal Assessment of Cellular Stress
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NFE2L2 (16396-1-AP) and LC3II (14600-1-AP) were purchased from proteintech. HO-1 (86806, CST), FTH1 (4393, CST), Gpx4 (PA5-102521, Invitrogen), NCOA4 (HC071-1F11, Sigma-Aldrich).
Esculetin, Cisplatin and Sorafenib (MCE, USA), T-AOC Assay, ROS Assay Kit, Crystal Violet Staining Solution, Lyso-Tracker Red (Solarbio, China), Mito-Tracker (Beyotime, China), DPPH free radical scavenging rate detection kit (Leagene, China), TranswellTM polycarbonate membrane cell culture inserts (Corning, Germany), Liperfluo, FerroOrange, Cell Counting Kit-8 (DOJINDO, Japan), GSH (reduced glutathione) determination kit (NJJCBIO).
Esculetin, Cisplatin and Sorafenib (MCE, USA), T-AOC Assay, ROS Assay Kit, Crystal Violet Staining Solution, Lyso-Tracker Red (Solarbio, China), Mito-Tracker (Beyotime, China), DPPH free radical scavenging rate detection kit (Leagene, China), TranswellTM polycarbonate membrane cell culture inserts (Corning, Germany), Liperfluo, FerroOrange, Cell Counting Kit-8 (DOJINDO, Japan), GSH (reduced glutathione) determination kit (NJJCBIO).
8
IM-CS-NPs/E7/ATP@ALG Nanovaccine Preparation
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Lactic acid-ethanolic acid (PLGA) copolymer 50/50 was obtained from Jinan Daigang Bioengineering Co; chitosan and PVA were purchased from Sigma; the Flow cytometry antibodies were provided by Biolegend, USA; LysoTracker Red were provided by Solarbio (L8010). DAPI Stain Solution (ab104139), Na+/K+-ATPase antibody (ab76020), HMGB1 antibody (ab79823), Donkey Anti-Rabbit IgG H&L antibody (ab150075) and Goat Anti-Rabbit IgG H&L antibody (ab150077) were purchased from Abcam; the antibodies for Calreticulin (27298–1-AP), HSP70 (10995–1-AP), HSP90 (13171–1-AP), HRP Goat Anti-Rabbit IgG antibody (SA00001-2) and ELISA kits (IL-1β, IL6, TNF-α and IFN-γ) were purchased from Proteintech (Scheme 1 ).![]()
Schematic illustration for the preparation of IM-CS-NPs/E7/ATP@ALG nanovaccine and its effects inducing antitumor immunity
9
Lysosomal Activity Monitoring Protocol
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LysoTracker Red (L8010, Solarbio Life Sciences) was diluted in DMEM at a ratio of 1:20,000 and incubated at 37°C for 2 hours in the dark. The area and intensity of red fluorescence indicated the lysosomal activity.
10
Multifunctional Vaccine Delivery System
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Stannous octoate (Sn(Oct)2), polyethyleneimine (PEI), ε-caprolactone, 4-nitrophenyl chloroformate (NPC), N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide (EDC), cysteamine hydrochloride, 2,2′-Dithiodipyridine, N-hydroxysuccinimide (NHS) were obtained from Aladdin (China). CpG ODN1826 and traut's reagent were obtained from Sangon Biotech (Shanghai). HOOC-PEG-NH2 were purchased from Ponsure Biological (Shanghai). Carboxyfluorescein succinimidyl ester (CFSE), OVA, OVA257−264, and OVA323−339 were purchased from Sigma-Aldrich (USA). LysoTracker Red and DAPI were purchased from Beijing Solarbio Science & Technology Co.,Ltd. The anti-SIINFEKL/H-2KbPE, anti-CD11c-BB700, anti-CD86-PE, anti-CD80-FITC, anti-CD3e-FITC, anti-CD4-APC and anti-CD8a-PerCP-Cy5.5 were obtained BD Biosciences (USA). IL-12p70, TNF-α and IFN-γ enzyme-linked immunosorbent assay (ELISA) kits were purchased from Invitrogen (USA). FITC—OVA was prepared according to the previous protocol.
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