Cells were seeded at 5×104 cells/well and were incubated with Rot (0.3 µM) with or without Re (5 µM) at 37°C for 24 h, washed twice with PBS, mixed with Rhodamine 123 (2 µM) (Beijing Solarbio Science & Technology Co., Ltd.; cat. no. R8030), and incubated at 37°C for 30 min in the dark. The cells were then centrifuged at 530 × g for 5 min at room temperature, resuspended in PBS, and analyzed using a flow cytometer (Amnis Corporation), and quantified using IDEAS software v6.1 (Amnis Corporation).
Rhodamine 123
Manufactured by Solarbio
Sourced in China
Rhodamine 123 is a fluorescent dye commonly used in various laboratory applications. It is a cationic dye that can be used to stain mitochondria in living cells, enabling the visualization and analysis of mitochondrial structure and function. Rhodamine 123 is known for its high quantum yield and photostability, making it a valuable tool in fluorescence microscopy and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Cell counting kit 8 cck 8, by MedChemExpress (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by CoWin Biotech (1 mentions)
β tubulin, by Merck Group (1 mentions)
Flag tag (flag), by Merck Group (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Dulbecco s modified eagle s medium dmem, by Corning (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Paraformaldehyde (pfa), by Solarbio (1 mentions)
Triton x 100, by Solarbio (1 mentions)
Paclitaxel, by Solarbio (1 mentions)
Adriamycin, by Solarbio (1 mentions)
Verapamil, by Solarbio (1 mentions)
Mitoxantrone, by Solarbio (1 mentions)
Goat anti rabbit igg h l secondary antibody, by Abcam (1 mentions)
Pgp glo assay system, by Promega (1 mentions)
Abcg2, by Abcam (1 mentions)
Anti p glycoprotein antibody, by Abcam (1 mentions)
Ko143, by MedChemExpress (1 mentions)
Goat anti mouse igg h l secondary antibody, by Abcam (1 mentions)
12 protocols using rhodamine 123
1
Mitochondrial Membrane Potential Assay
2
Multidrug-Resistance Modulation Protocols
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Rutaecarpine, paclitaxel, adriamycin, colchicine, and cisplatin were purchased from Chengdu Alfa Biotechnology Co. Ltd. (Chengdu, China). Rhodamine 123, 4′,6-diamidino-2-phenylindole (DAPI), Triton X-100, 4% paraformaldehyde, and enhanced DAB reagent kit were purchased from Solarbio Science & Technology Co. Ltd. (Beijing, China). Tariquidar (ABCB1 inhibitor) was obtained from Beyotime Biotechnology (Shanghai, China). Cell counting kit-8 (CCK8) was purchased from Med Chem Express (MCE). Annexin V-FITC/PI apoptosis detection kit was purchased from Beijing ComWin Biotech Co. Ltd. (CWBIO) (Beijing, China). Fetal bovine serum (FBS) and Dulbecco’s modified Eagle’s Medium (DMEM) were purchased from Corning Incorporated (Corning, New York, NY, USA). The antibodies used for WB, IFC, Co-IP, and IH were raised against the following proteins: ABCB1 (Zen Bioscience, Chengdu, China), β-tubulin (Sigma-Aldrich, St. Louis, MO, USA), Flag (Sigma-Aldrich St. Louis, MO, USA), Myc (Santa Cruz, CA, USA), HA (Santa Cruz, CA, USA), β-actin (Santa Cruz, CA, USA), MARCH8 (BBI Life Science, Shanghai, China), horseradish peroxidase (HRP)-conjugated secondary antibodies to mouse (Beyotime, Shanghai, China), or rabbit (Beyotime, Shanghai, China).
3
Efficient Multidrug Resistance Inhibition
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The σ2 receptor ligand A011 was prepared as previously reported (Feng et al., 2019 (link)). KO143 was purchased from MedChemExpress (Monmouth Junction, NJ, United States). Cisplatin (DDP), adriamycin (ADR), paclitaxel, verapamil, mitoxantrone and rhodamine 123 (Rh123) were acquired from Solarbio (Beijing, China). Cell Counting Kit-8 (CCK-8) was bought from Dojindo Laboratories (Japan). Pgp-Glo™ Assay Systems were purchased from Promega (Madison, USA). Anti-P Glycoprotein antibody, ABCG2, GAPDH, Goat Anti-Rabbit IgG H&L Secondary Antibody (Alexa Fluor 488), Goat Anti-Rabbit IgG H&L Secondary Antibody and Goat Anti-Mouse IgG H&L Secondary Antibody were purchased from Abcam (Cambridge, United Kingdom). SlowFadeTM Gold antifade reagent was purchased from Thermo Fisher (MA, United States).
4
Synthesis and Characterization of PEGylated Gold Nanoparticles
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Gold chloride solution (HAuCl4) and tetraethyl orthosilicate (TEOS) were purchased from Aladdin Industrial Co., Ltd. (Shanghai, China); polyethylene glycol (NH2-PEG-COOH; molecular weight 3070) from Shanghai ZZBIO Co., Ltd. (Shanghai, China); fetal bovine serum (FBS) and Roswell Park Memorial Institute (RPMI) 1640 culture medium from Gibco (Invitrogen, Carlsbad, CA, USA); HA, acridine orange (AO), fluorescein diacetate (FDA), propidium iodide (PI), Hoechst 33258, 4ʹ,6-diamidino-2-phenylindole (DAPI), 3-(4,5-dimethylthiazlo-2-diphenyl-tetrazolium) bromide (MTT), Rhodamine B isothiocyanate (RBITC), PTX, and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA); an annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit from Keygen Biotech Co., Ltd. (Nanjing, China); Rhodamine 123 and 2ʹ-7ʹ dichlorofluorescin diacetate (DCFH-DA) from Solarbio Technology Co., Ltd. (Beijing, China). Monoclonal mouse anti-P-gp antibody, Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG were obtained from Proteintech Group, Inc. (Chicago, IL, USA); other primary antibodies were purchased from Sigma-Aldrich (St. Louis, MO, USA). Other reagents and chemicals were of analytical grade unless otherwise stated and were purchased from local commercial suppliers. Deionized (DI) water (Milli-Q, Millipore, Bedford, MA, USA) was used to prepare aqueous solutions.
5
Fetal Bovine Serum and CHLORIN E6 in ROS Assay
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Certified fetal bovine serum (04-001-1ACS, VivaCell, Shanghai, China), DMEM medium (SH30022.01B, HyClone, Beijing, China), CHLORIN E6 (C829662, MACKIN, Shanghai, China), Rhodamine123 (R8030, Solarbio, Beijing, China), ROS Assay Kit (S0033S, Beyotime, Shanghai, China), Annexin-V-iFluor488/PI (CM001-100D, CHAMOT, Shanghai, China), γ-H2AX (ET1705-97, HUABIO, Hangzhou, China), p-ATM (ET1705-50, HUABIO, Hangzhou, China), GADPH (PMK053C, BioPM, Wuhan, China), CD 41 (bs-2636R, Bioss, Beijing, China), CD62p (ab83583, Abcam, Cambridge, UK), Annexin-V (DF6580, Affinity, USA), and D-luciferin, potassium salt (luc001, Sciencelight, Shanghai, China) were used in our study.
6
Mitochondrial Membrane Potential and ROS Assay
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To determine the MMP, genipin-treated HCT116 cells were stained with 10 µM Rhodamine 123 (Solarbio) for 30 min and evaluated by fluorescence microscopy. To determine ROS generation, genipin-treated HCT116 cells were stained with 20 µM DCFH-DA (Solarbio) for 30 min and evaluated by flow cytometry.
7
Advanced Cell Apoptosis Assays
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KM (lot NO., 17020911, purity > 98%) was purchased from Manster Biotechnology Co., Ltd. (Cheng du, China). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from KeyGen Biotechnology Co., Ltd. (Nanjing, China). Bovine serum albumin (BSA), D-glucose and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Penicillin, streptomycin, rhodamine 123, hoechst 33258, reactive oxygen species assay kit and Bicinchoninicacid (BCA) protein concentration determination kit were purchased from Solarbio Technology Co., Ltd. (Beijing, China). Anti-rat RAGE (ab65965), Bax (ab32503), Bcl-xL (ab32370), Cytochrome C (ab133504), Cleaved-caspase-3 (ab13847), Cleaved-caspase-9 (ab25758), Cleaved-PARP (ab32138), TGF-β1 (ab92486), Collagen IV (ab6586) and β-actin (ab8227) were purchased from Abcam Biotechnology Co., Ltd. (Shanghai, China). The annexin V and PI apoptosis kits were purchased from Invitrogen Thermo Fisher Scientific (Carlsbad, CA, USA). The AGEs Enzyme-linked Immuno Sorbent Assay (ELISA) kit was obtained from R&D Systems (Minneapolis, MN, USA). The superoxide dismutase (SOD) kit and Malondialdehyde (MDA) kit were purchased from Jiancheng Bioengineering Institute (Nanjing, China).
8
Rhodamine 123 Staining of Yeast Cells
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Yeast cells were mixed with Rhodamine 123 (2 µM; cat # R8030, Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) and incubated in darkness at 37 °C for 30 min. After centrifugation at 530× g for 5 min, a flow cytometer (Amnis Corporation, Seattle, WA, USA) was used for detection, and IDEAS software v6.1 was used for quantitative analysis.
9
DNA Condensation and Nuclear Fragmentation
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rhodamine123 assays were performed to determine the DNA condensation and nuclear fragmentation. After the required treatment, cells were incubated with the 10 μmol/L rhodamine123 (Solarbio, R8030) at 37 °C for 1.5 h in the dark. The fluorescence intensity of harvested cells was detected using an Olympus IX-73 microscope.
10
Mitochondrial Activity Assay of BMSCs
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BMSCs-GSSM-gels were washed twice with PBS. Rhodamine 123 (#R8030, Solarbio, China) was applied at a working concentration of 50 μL/mL and incubated at 37°C for 5 minutes, with the resultant staining observed under a fluorescence microscope.
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