3 3 dihexyloxacarbocyanine iodide dioc6 3

Manufactured by Thermo Fisher Scientific

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3,3′-dihexyloxacarbocyanine iodide [DiOC6(3)], is a lipophilic fluorescent dye. It can be used to stain mitochondria in live cells.

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Lab products found in correlation

2 7 dichlorodihydrofluorescein diacetate h2dcfda, by Thermo Fisher Scientific (4 mentions) Z vad fmk, by Merck Group (3 mentions) Propidium iodide, by Merck Group (2 mentions) 2 7 dichlorofluorescin diacetate dcfda, by Merck Group (2 mentions) Caspase 3, by R&D Systems (2 mentions) Dulbecco s modified eagle s medium dmem, by Merck Group (2 mentions) Quercetin, by Merck Group (2 mentions) Propidium iodide, by Thermo Fisher Scientific (2 mentions) Entinostat, by Selleck Chemicals (1 mentions) Irinotecan hydrochloride cpt 11, by Merck Group (1 mentions) Navitoclax, by Selleck Chemicals (1 mentions) Z vad fmk, by Selleck Chemicals (1 mentions) Chloramphenicol, by Carl Roth (1 mentions) Rotenone, by Abcam (1 mentions) Irinotecan hydrochloride, by Merck Group (1 mentions) Doxorubicin, by Enzo Life Sciences (1 mentions) Carbonyl cyanide 4 trifluoromethoxy phenylhydrazone fccp, by Abcam (1 mentions) Antimycin a, by Merck Group (1 mentions) Oligomycin, by Abcam (1 mentions) Penicillin g, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

DiOC6 (109 citations) DiOC6(3) (74 citations) 3,3′-dihexyloxacarbocyanine iodide (DiOC6) (14 citations) 3,3′-Dihexyloxacarbocyanine Iodide (5 citations) 3,3-Dihexyloxacarbocyanine (DiOC6, (2 citations)

17 protocols using 3 3 dihexyloxacarbocyanine iodide dioc6 3

Apoptosis Induction Assay Protocol

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Irinotecan hydrochloride (CPT‐11) and propidium iodide (PI) were purchased from Sigma‐Aldrich (Deisenhofen, Germany). Entinostat (MS‐275), navitoclax (ABT‐263), and z‐VAD‐FMK (zVAD) were purchased from Selleck Chemicals (Houston, TX, USA). 3,3′‐dihexyloxacarbocyanine iodide (DiOC₆(3)) was purchased from Thermo Fisher Scientific. CPI‐1612 [24 (link)] was kindly provided from M. Trojer (Constellation Pharmaceuticals, Cambridge, MA, USA). DMSO was used as treatment control.
All other chemicals were bought from Carl Roth GmbH & Co KG (Karlsruhe, Germany). All buffers and solutions were prepared with double‐distilled water (ddH₂O).

Marx C., Sonnemann J., Beyer M., Maddocks O.D., Lilla S., Hauzenberger I., Piée‐Staffa A., Siniuk K., Nunna S., Marx‐Blümel L., Westermann M., Wagner T., Meyer F.B., Thierbach R., Mullins C.S., Kdimati S., Linnebacher M., Neri F., Heinzel T., Wang Z, & Krämer O.H. (2021). Mechanistic insights into p53‐regulated cytotoxicity of combined entinostat and irinotecan against colorectal cancer cells. Molecular Oncology, 15(12), 3404-3429.

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Mitochondrial Dysfunction and Oxidative Stress Assays

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Irinotecan hydrochloride, 2-thenoyltrifluoroacetone (TTFA), hydroxyurea (HU), 2′,7′-dichlorofluorescin diacetate (DCFDA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2-deoxy-d-glucose (2-DG), crystal violet, and antimycin A were purchased from Sigma Aldrich (Deisenhofen, Germany). Doxorubicin was purchased from Enzo Life Science (Lörrach, Germany). Chloramphenicol was purchased from Carl Roth GmbH (Karlsruhe, Germany). Rotenone, carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP), and oligomycin were purchased from Abcam (Cambridge, UK). 3,3′-Dihexyloxacarbocyanine iodide (DiOC₆(3)) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). DMSO was used as a treatment control.

Marx C., Sonnemann J., Maddocks O.D., Marx-Blümel L., Beyer M., Hoelzer D., Thierbach R., Maletzki C., Linnebacher M., Heinzel T, & Krämer O.H. (2022). Global metabolic alterations in colorectal cancer cells during irinotecan-induced DNA replication stress. Cancer & Metabolism, 10, 10.

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Apoptosis Induction in Cancer Cells

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Cisplatin, propidium iodide (PI) and thiazolyl blue tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Trypsin-EDTA was purchased from BioConcept (Allschwil/BL, Switzerland). Fetal bovine serum (FBS), L-glutamine, penicillin G, 2’, 7’-dichlorodihydrofluorescein diacetate (H₂DCFDA) and 3, 3-dihexyloxa-carbocyanine iodide [DiOC₆(3)] were obtained from Thermo Fisher Scientific (Carlsbad, CA, USA). Caspase-3 and caspase-9 activity assay kits were purchased from R&D Systems Inc. (Minneapolis, MN, USA). The primary antibodies against Bcl-2, Bax, cytochrome c, Apaf-1, AIF, p21, cyclin A, cyclin D, cyclin E, CDK 2, β-actin and the goat anti-rabbit or anti-mouse IgG-horseradish peroxidase (HRP) secondary antibodies were purchased from GeneTex, (Hsinchu, Taiwan). Pan-caspase inhibitor (z-VAD-fmk) and enhanced chemiluminescence (ECL) detection kit (Immobilon Western Chemiluminescent HRP Substrate) were purchased from Merck Millipore (Billerica, MA, USA). YC-1 was designed and synthesized as detailed in the previous study [21 (link)].

Lee M.R., Lin C., Lu C.C., Kuo S.C., Tsao J.W., Juan Y.N., Chiu H.Y., Lee F.Y., Yang J.S, & Tsai F.J. (2017). YC-1 induces G0/G1 phase arrest and mitochondria-dependent apoptosis in cisplatin-resistant human oral cancer CAR cells. BioMedicine, 7(2), 12.

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Mitochondrial Membrane Potential Assay

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3,3′-Dihexyloxacarbocyanine iodide (DiOC6(3)) was purchased from Thermofisher Scientific and applied to monitor mitochondrial membrane potential according to the manufacturer’s instructions (San Jose, CA, USA). Cells were seeded into six-well plates and treated with vehicle or AA for 24 h. Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was also applied as a positive control, except that cells were harvested after brief exposure, i.e., 6 h. Cells were trypsinized and washed with PBS two times, followed by DiOC6(3) staining for 20 min in 37 °C water bath. Then cells were washed with PBS for an additional three times, re-suspended with HBSS, and submitted to flow cytometry for analysis (Becton-Dickinson, Franklin Lakes, NJ, USA).

Yi H., Wang K., Du B., He L., HO H., Qiu M., Zou Y., Li Q., Jin J., Zhan Y., Zhao Z, & Liu X. (2018). Aleuritolic Acid Impaired Autophagic Flux and Induced Apoptosis in Hepatocellular Carcinoma HepG2 Cells. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(6), 1338.

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PBMC Viability Assessment Assay

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Peripheral blood mononuclear cells (PBMCs) were purified by standard density gradient centrifugation using Ficoll-Hypaque (Atom Reactiva, Barcelona, Spain) and used immediately for ex-vivo SCD assays. B-cell death was evaluated by culturing PBMC (1 × 10 6 cells/ml) in Roswell Park Memorial Institute 1640 medium (RPMI-1640) supplemented with 10% of FBS (R10 medium; ThermoFisher Scientific) for 24 h. Then, PBMCs were harvested and incubated with 40 nmol/l of the potentiometric mitochondrial probe 3,3'-dihexyloxacarbocyanine Iodide (DIOC 6 (3)) (ThermoFisher Scientific) in R10 for 1 h at 37°C/5% CO 2 . After that, cells were stained with anti-CD19 APC/Cy7, anti-IgD PE, anti-CD38 PerCP-Cy5.5, anti-CD5 APC (BD Biosciences), and anti-CD27 PE/Cy7 (eBiosciences). Finally, 0.3 μm of Sytox Blue (ThermoFisher Scientific) was added and incubated for 10 min. Cells were acquired in a LSR-II flow cytometer (BD Biosciences). Live cells were identified as DIOC 6 (3) bright/Sytox Blue negative using FlowJo software (Tree Star Inc., Ashland, Oregon, USA).

Carrillo J., Negredo E., Puig J., Molinos-Albert L.M., Rodríguez de la Concepción M.L., Curriu M., Massanella M., Navarro J., Crespo M., Viñets E., Millá F., Clotet B, & Blanco J. (2018). Memory B cell dysregulation in HIV-1-infected individuals. AIDS (London, England), 32(2).

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Apoptosis and Oxidative Stress Assays

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We obtained an In Situ Cell Death Detection Kit (Fluorescein), thiazolyl blue tetrazolium bromide (MTT), indomethacin, quercetin, Dulbecco’s modified Eagle’s medium (DMEM), DNase, and other reagents and chemicals from Sigma–Aldrich, Merck KGaA (Darmstadt, Germany). We obtained Muse Caspase-3/9 Assay Kits and the pan-caspase inhibitor Z-VAD-FMK from Millipore, Merck KGaA (Darmstadt, Germany). We obtained dihydroethidium, 2′,7′-dichlorodihydrofluorescein diacetate (H₂DCFDA), 3,3′-dihexyloxacarbocyanine iodide [DiOC6(3)], and MitoSOX from Molecular Probes, Thermo Fisher Scientific (Waltham, MA, USA). We purchased L-glutamine, fetal bovine serum (FBS), trypsin-EDTA, and penicillin/streptomycin from HyClone, GE Healthcare Life Sciences (Logan, UT, USA).

Chen C., Yang J.S., Lu C.C., Wu Y.T, & Chen F.A. (2021). Effect of Quercetin on Injury to Indomethacin-Treated Human Embryonic Kidney 293 Cells. Life, 11(11), 1134.

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Dual-Targeting Cytotoxicity Evaluation

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HEPG2 and HUH7 cells were seeded at 1.5 × 10⁵ cells/well and 1 × 10⁵ cells/well, respectively, 24 h prior to treatment. Groups were treated with 0.0, 0.05 and 0.1 µM of NV651 in combination with 0, 5 and 10 µM of Cisplatin for 72 h. The supernatant and trypsinized cells were collected for analysis. Viability was quantified using propidium iodide (PI) (Thermo Fisher Scientific, Waltham, MA, USA) at a concentration of 1 μg/mL to evaluate the plasma membrane integrity and mitochondrial membrane potential with 3,3′-dihexyloxacarbocyanine iodide (DiOC(6)3) (Molecular Probes–Invitrogen, Waltham, MA, USA) staining at a concentration of 40 nM.

Simón Serrano S., Tavecchio M., Mallik J., Grönberg A., Elmér E., Kifagi C., Gallay P., Hansson M.J, & Massoumi R. (2022). Synergistic Effects of Sanglifehrin-Based Cyclophilin Inhibitor NV651 with Cisplatin in Hepatocellular Carcinoma. Cancers, 14(19), 4553.

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Microbial Cell Viability and Enzyme Assays

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BacTiter-Glo^™ Microbial Cell Viability and NADP/NADPH-Glo^™ Assay Kits were from Promega (Madison, WI, USA). Z-Ala-Arg-Arg-7-amino-4-methylocoumarin (Z-ARR-AMC) was from Merck Millipore (Darmstadt, Germany). Suc-Leu-Leu-Val-Tyr-7-amino-4-methylocoumarin (Suc-LLVY-AMC) and Z-Leu-Leu-Glu-7-amino-4-methylocoumarin (Z-LLE-AMC) were from Enzo Life Sciences (Farmingdale, NY, USA). FUN^® 1 Cell Stain, rhodamine B hexyl ester and 3,3′-Dihexyloxacarbocyanine Iodide (DiOC6(3)) were from Molecular Probes (Eugene, OR, USA). Coomassie Protein Assay Reagent (Thermo Scientific, Rockford, IL, USA) All other reagents were purchased from Sigma-Aldrich (Poznan, Poland). Components of culture media were from BD Difco (Becton Dickinson and Company, Spark, MD, USA) except for glucose (POCH, Gliwice, Poland).

Maslanka R, & Zadrag-Tecza R. (2020). Reproductive Potential of Yeast Cells Depends on Overall Action of Interconnected Changes in Central Carbon Metabolism, Cellular Biosynthetic Capacity, and Proteostasis. International Journal of Molecular Sciences, 21(19), 7313.

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Cytotoxicity and Apoptosis Assay Protocol

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Quercetin, dexamethasone, thiazolyl blue tetrazolium bromide (MTT), the In Situ Cell Death Detection Kit (Fluorescein), and other chemicals and reagents were purchased from Sigma–Aldrich, Merck KGaA (Darmstadt, Germany), unless otherwise stated. All primary antibodies and anti-mouse and antirabbit immunoglobulin (Ig) G horseradish peroxidase (HRP)-linked secondary antibodies were purchased from GeneTex (Hsinchu, Taiwan). Z-VAD-FMK (a pan-caspase inhibitor) and Muse Caspase-3/9 assay kits were obtained from Millipore, Merck KGaA (Darmstadt, Germany). 2′, 7′-Dichlorodihydrofluorescein diacetate (H2DCFDA) and 3, 3′-dihexyloxacarbocyanine iodide [DiOC6(3)] were obtained from Molecular Probes, Thermo Fisher Scientific (Waltham, MA, USA). Dulbecco’s modified Eagle’s medium (DMEM) was purchased from Sigma–Aldrich (Lenexa, KS, USA). Fetal bovine serum (FBS), L-glutamine, penicillin/streptomycin, and trypsin-EDTA were purchased from HyClone, GE Healthcare Life Sciences (Logan, UT, USA).

Chen C., Yang J.S., Lu C.C., Chiu Y.J., Chen H.C., Chung M.I., Wu Y.T, & Chen F.A. (2020). Effect of Quercetin on Dexamethasone-Induced C2C12 Skeletal Muscle Cell Injury. Molecules, 25(14), 3267.

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Simultaneous Assessment of Cell Membrane Integrity and Mitochondrial Potential

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For the simultaneous quantification of plasma membrane integrity and mitochondrial transmembrane potential (Δψm), both adherent and non-adherent cells were collected, washed, and costained for 30 min at 37°C in 300 µL of culture medium containing 1 µg/mL propidium iodide (PI) which only incorporates into dead cells exhibiting plasma membrane rupture, and 40 nmol/L 3,3’-dihexyloxacarbocyanine iodide DiOC₆(3), a mitochondrial transmembrane potential sensitive dye (Molecular Probes-Invitrogen).18–20 (link) Cytofluorometric acquisitions were carried out on a Miltenyi cytofluorometer (MACSQuant Analyzer V.10), and analyses were performed by using the FlowJo software V.10.4.2 (TreeStar) on gating on events exhibiting normal forward scatter and side scatter parameters.

Juncheng P., Joseph A., Lafarge A., Martins I., Obrist F., Pol J., Saavedra E., Li S., Sauvat A., Cerrato G., Lévesque S., Leduc M., Kepp O., Durand S., Aprahamian F., Nirmalathansan N., Michels J., Kroemer G, & Castedo M. (2022). Cancer cell-autonomous overactivation of PARP1 compromises immunosurveillance in non-small cell lung cancer. Journal for Immunotherapy of Cancer, 10(6), e004280.

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