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Mitoprobe diic1 5 assay kit

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The MitoProbe DiIC1(5) Assay Kit is a fluorescent dye-based tool used to measure mitochondrial membrane potential in cells. The kit provides the DiIC1(5) dye, a cationic carbocyanine compound that accumulates in active mitochondria in proportion to their membrane potential.

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Lab products found in correlation

Tmre mitochondrial membrane potential assay kit, by Abcam (1 mentions) Luciferase assay system, by Promega (1 mentions) Bafilomycin a1 bafa1, by Merck Group (1 mentions) Anti parkin antibody, by Santa Cruz Biotechnology (1 mentions) Z vad ome fmk, by Santa Cruz Biotechnology (1 mentions) Annexin 5 fitc, by Thermo Fisher Scientific (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Mitosox, by Thermo Fisher Scientific (1 mentions) N acetyl cysteine (nac), by Merck Group (1 mentions) Anti p62 antibody, by Merck Group (1 mentions)

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DiIC1(5) (9 citations) MitoProbe DiIC1(5) kit (5 citations) MitoProbe DiIC1(5) (5 citations)

6 protocols using mitoprobe diic1 5 assay kit

1

Measuring Mitochondrial Membrane Potential

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The MitoProbe™ DiIC1(5) Assay Kit (M34151, Thermo Fisher Scientific) and TMRE-Mitochondrial Membrane Potential Assay Kit (ab113852, Abcam) were used to determine the mitochondrial membrane potential (MMP). We first treated the cells with magnolol, then measured the MMP by using fluorescence microscopy and/or flow cytometry according to the manufacturer's protocol. In each independent experiment, the fluorescence intensity of the DMSO control groups was normalized to 100%, and the fluorescence intensity of magnolol-treated groups was subsequently compared with control groups to determine the remaining fluorescence intensity. Three independent experiments were performed to get the statistical results.
Tang Y., Wang L., Yi T., Xu J., Wang J., Qin J.J., Chen Q., Yip K.M., Pan Y., Hong P., Lu Y., Shen H.M, & Chen H.B. (2021). Synergistic effects of autophagy/mitophagy inhibitors and magnolol promote apoptosis and antitumor efficacy. Acta Pharmaceutica Sinica. B, 11(12), 3966-3982.
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2

Mitochondrial Membrane Potential Assay

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Mitochondrial membrane potential was assessed using the MitoProbe DiIC1(5) Assay Kit (Thermo Fisher Scientific). THP-1 cells were cultured with or without PMPs for 24 h before 5 × 105 cells in 1 mL IMDM + 10% FBS were stained with DiIC1(5) using carbonyl cyanide 3-chlorophenylhydrazone-treated cells as a correction for background signal and incubated for 30 min following the manufacturer’s instructions. After doublet discrimination, MFI (mean fluorescence intensity) values of gated cells were compared using the APC channel on the flow cytometer. See Figure S3A and B for gating strategy. At least 30,000 gated cells were collected.
Cacic D., Reikvam H., Nordgård O., Meyer P, & Hervig T. (2021). Platelet Microparticles Protect Acute Myelogenous Leukemia Cells against Daunorubicin-Induced Apoptosis. Cancers, 13(8), 1870.
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3

Metastatic Breast Cancer Modeling

Cited 2 times
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All animal studies were carried out in accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Institutional Animal Care and Use Committee of the University of Southern California. MCF-7 and MDA-MB-231 breast cancer cells (ATCC), stably transduced with a GFP/luciferase construct62 (link),63 (link), were stained with TMRM or DiIC1(5) (MitoProbe DiIC1(5) Assay Kit, Invitrogen) as a marker of mitochondrial membrane potential and sorted on the BD FACSAria I cell sorter (Flow Cytometry Core Facility, USC). Sorted cells were inoculated into NSG (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ) mice through tail vein injections (300,000 cells per mouse for MCF-7 and 100,000 cells per mouse for MDA-MB-231). For the MCF-7 study, mice were implanted with 17β-estradiol 90-day release pellets (0.36 mg/pellet), using a sterile trochar 24 hours prior to the injections. After five weeks (for MCF-7 cells) or four weeks (for MDA-MB-231 cells), the animals were euthanized and their lungs were harvested, ground, and assayed for metastatic tumor content using a Luciferase Assay System (Promega)17 .
Begum H.M., Ta H.P., Zhou H., Ando Y., Kang D., Nemes K., Mariano C.F., Hao J., Yu M, & Shen K. (2019). Spatial Regulation of Mitochondrial Heterogeneity by Stromal Confinement in Micropatterned Tumor Models. Scientific Reports, 9, 11187.
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4

Mitochondrial Membrane Potential Analysis

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Cells were treated as indicated in the figure legends. (Ⅰ) After treatment, cell pellets were collected and resuspended in MitoProbe™ DiIC1(5) Assay Kit (Invitrogen, M34151) with a final concentration of 25 nM. After incubation at 37 °C, 5% CO2 for 15 min, cells were centrifuged and resuspended in 500 μL ice-cold PBS and subsequently analyzed on a flow cytometer with 633 nm excitation to determine the fluorescence intensity. (Ⅱ) After treatment, cell pellets were stained with Tetramethylrhodamine, Ethyl Ester, Perchlorate (TMRE) (Abcam, ab113852) with a final concentration of 200 nM for 20 min. Cells were collected and resuspended in 500 μL ice-cold PBS and subsequently analyzed on a flow cytometer to determine the fluorescence intensity.
Xu J., Wang L., Zhang L., Zheng F., Wang F., Leng J., Wang K., Héroux P., Shen H.M., Wu Y, & Xia D. (2020). Mono-2-ethylhexyl phthalate drives progression of PINK1-parkin-mediated mitophagy via increasing mitochondrial ROS to exacerbate cytotoxicity. Redox Biology, 38, 101776.
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5

Mitochondrial Dynamics and Apoptosis Assays

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Our experiments used the following reagents: Mono-2-ethylhexyl phthalate (MEHP, Sigma 796,832), Di-2-ethylhexyl phthalate (DEHP, Sigma 80,030), Carbonyl cyanide 3-chlorophenylhydrazone (CCCP, Sigma C2759), N-Acetyl Cysteine (NAC, Merck), Z-VAD (OMe)-FMK (Santa Cruz, CAS 187389-52-2), Bafilomycin A1 (BafA1, Sigma B1793), MitoSOX™ (Invitrogen, M36008), MitoProbe™ DiIC1(5) Assay Kit (Invitrogen, M34151), Tetramethylrhodamine, Ethyl Ester, Perchlorate (TMRE) (Abcam, ab113852), Propidium iodide (PI, Sigma P4170), Annexin V FITC (Invitrogen, V13242). We used the following antibodies: anti-phospho-DRP1 (Ser616) antibody (CST, 3455), anti-phospho-DRP1 (Ser637) antibody (CST, 4867), anti-DRP1 (D6C7) antibody (CST, 8570), anti-HSP60 (D6F1) antibody (CST, 12,165), anti-Parkin antibody (Santa Cruz, SC32282), anti-PINK1 (D8G3) antibody (CST, 6946), anti-Tim23 (H-8) antibody (Santa Cruz, SC514463), anti-Tom20 (FL-145) antibody (Santa Cruz, SC11415), anti-COXIV (3E11) antibody (CST, 4850), anti-Mitofusin-1 (D6E2S) antibody (CST, 14,739), anti-Mitofusin-2 (D1E9) antibody (CST, 11,925), anti-VDAC antibody (CST, 4866), anti-microtubule-associated protein 1 light chain 3/LC3 antibody (Sigma, L7543), anti-p62 antibody (Sigma, SAB1406748), anti-phospho-ubiquitin antibody (Merck Millipore, ABS1513-I), anti-ubiquitin (P4D1) antibody (Santa Cruz, SC8017), anti-β-actin antibody (Sigma, A5441).
Xu J., Wang L., Zhang L., Zheng F., Wang F., Leng J., Wang K., Héroux P., Shen H.M., Wu Y, & Xia D. (2020). Mono-2-ethylhexyl phthalate drives progression of PINK1-parkin-mediated mitophagy via increasing mitochondrial ROS to exacerbate cytotoxicity. Redox Biology, 38, 101776.
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6

Mitochondrial Membrane Potential Assessment

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We assess the mitochondrial membrane potential using MitoProbe DiIC1(5) Assay Kit (Invitrogen, M34151). The mitochondrial membrane potential was measured using DiIC(5) staining according to the manufacturer’s instructions. CCCP treatment was used as a positive control for depolarization.
Huang Y., Shao M., Teng X., Si X., Wu L., Jiang P., Liu L., Cai B., Wang X., Han Y., Feng Y., Liu K., Zhang Z., Cui J., Zhang M., Hu Y., Qian P, & Huang H. (2024). Inhibition of CD38 enzymatic activity enhances CAR-T cell immune-therapeutic efficacy by repressing glycolytic metabolism. Cell Reports Medicine, 5(2), 101400.
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