Trolox

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Trolox is a water-soluble vitamin E analog that functions as an antioxidant. It is commonly used as a reference compound in various biological and biochemical assays.

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Lab products found in correlation

15 protocols using trolox

Cell Culture and Metabolic Labeling

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U2OS cells (American Tissue and Cell Collection, HTB-96) were a kind gift of Dr. John Hogenesh (University of Pennsylvania) NIH-3T3 (CRL-1658), MDCK (CCL-34) and 293T cells (CRL-3216) were directly obtained from ATCC. All cell lines were cultured in 10% Dulbecco’s modified Eagle medium supplemented to 10% decomplemented fetal bovine serum at 37°C, 5% CO₂, 21% O₂ and 100% humidity unless otherwise indicated. For aminoacid depletion experiments, dialyzed FBS was used and L-glutamine was supplemented as indicated. Cell lines were maintained and passaged according to ATCC recommended procedures. Pharmacological agents was as follows: Purvalanol-A (Sigma P4484), Z-VAD-FMK (EMD Millipore 627610), actinomycin D (Sigma A9415), cycloheximide (Sigma C7698), NAC (Sigma A9165), Trolox (Santa Cruz Biotech sc-200810) and CCCP (Sigma C2759) were added to cell cultures 12 h after cell seeding at the concentrations indicated and maintained until analysis, changing the medium every 24 hours. Ethynyl-uridine (Life Technologies E10345) and homopropargyl-glycine (Life Technologies C10186) were added at 5 and 50 μM respectively, two hours prior to incorporation analysis using the Click-iT alkyne detection kit (Life Technologies C10330).

Penzo-Méndez A.I., Chen Y.J., Li J., Witze E.S, & Stanger B.Z. (2015). Spontaneous Cell Competition in Immortalized Mammalian Cell Lines. PLoS ONE, 10(7), e0132437.

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Lipid Formulation Development and Characterization

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1,2-di-palmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Lipoid P-100 phosphatidylcholine (>97%) from soybean non-(GMO) was kindly supplied by Lipoid GmbH (Ludwigshafen, Germany). Cholesterol, curcumin (cur ≥ 65%), α-tocopherol (α-toc ≥ 96%), malondialdehyde (MDA), and 1,1,3,3-tetraethoxypropane (TEP) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Purified water was obtained from an ultrapure water system (Thermo Scientific Barnstead MicroPure ST, Langenselbold, Germany). HPLC grade chloroform, HPLC grade acetonitrile, HPLC grade methanol, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), 1, 1 diphenyl 2-picryl-hydrazyl (DPPH), thiobarbituric acid (TBA), and Triton X-100 were purchased from Merck. Trolox was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Dulbecco’s Modified Eagle Medium (DMEM), DMEM without phenol red, fetal bovine serum, penicillin, streptomycin, fetal bovine serum (FBS), and Trypsin-EDTA were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Peptone, Yeast Extract–Peptone–Dextrose Broth, and Sabouraud Dextrose Agar were obtained from Becton, Dickinson and Company (Sparks, MD, USA).

Vergara D., López O., Sanhueza C., Chávez-Aravena C., Villagra J., Bustamante M, & Acevedo F. (2023). Co-Encapsulation of Curcumin and α-Tocopherol in Bicosome Systems: Physicochemical Properties and Biological Activity. Pharmaceutics, 15(7), 1912.

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Autophagy and Oxidative Stress Regulation

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Rabbit anti-LC3B (L7543), mouse anti-βActin (A5441), N-Acetyl-L-cysteine (A9165) were from Sigma. Mouse anti-p62 (D-3, sc-28359), mouse anti-Lamp1 (H4A3, sc-20011), mouse anti-HIF-1α (28b, sc-13515), rabbit anti-Cathepsin B (sc-6493), mouse anti-Galectin 3 (sc-32790), DL-α-Tocopherol (sc-294383A), H-Leu-Leu-OMe Hydrochloride (sc-285992A), Trolox (sc-200810), Ferrocene (sc-353607), Deferoxamine mesylate (sc-203331) were from Santa Cruz Biotechnology. Rabbit anti-PARP (9542S) were from Cell Signaling. Acridine orange (A-3568) was from Life Technologies. Bafilomycin A1 (1334) was from Tocris. Chloroquine (C6628), Wortmannin (W1628), Ferrostatin-1 (SML0583), Necrostatin-1 (N9037), Cisplatin (P4394), Docetaxel (01885), Doxorubicin (D1515), Paclitaxel (T7402) were from Sigma. Phenethyl isothiocyanate (PEITC, 253731) was from Aldrich. E-64-D (BML-PI107), Pepstatin A (ALX-260-085) were from Enzo Life Sciences. Z-VAD-FMK (A1902) was from APExBIO. Ferroquine and amino-ferrocene were provided by Prof. Christophe Biot, University Lille1, France.

Kondratskyi A., Kondratska K., Vanden Abeele F., Gordienko D., Dubois C., Toillon R.A., Slomianny C., Lemière S., Delcourt P., Dewailly E., Skryma R., Biot C, & Prevarskaya N. (2017). Ferroquine, the next generation antimalarial drug, has antitumor activity. Scientific Reports, 7, 15896.

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Live Cell Imaging of Erythrocyte Invasion

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The sample was imaged in phenol red-free complete medium. For calcium experiments, the imaging medium was supplemented with 5 mM sodium pyruvate, 10 µM Trolox (Santa Cruz 53188-07-1) and 0.25 mM CaCl₂. For experiments involving inhibitors, either 100 µg/mL R1 peptide (China Peptides), 1 µg/mL cytochalasin D (Sigma Aldrich C8273), or 10 µM TRAM34 (Sigma Aldrich T6700) were added to the imaging medium. For confocal microscopy, 200 µL of imaging medium and 30 µL of stained erythrocytes per well were loaded to eight-well plate (Ibidi 80826). Then, 5–10 µL of stained schizonts were added to the stained erythrocytes in the first well right before imaging. For LLSM, an acid-washed 5 mm round glass coverslip (Warner Instruments CS-5R) was attached to the bottom of each well before loading 200 µL of phenol red-free RPMI-HEPES and 30 µL of stained erythrocytes to the well. Then, 5–10 µL of stained schizonts were gently added to the top of the coverslip and left to settle on the coverslip for 15 minutes. A tweezer was used to attach the coverslip to the sample stage, then the coverslip was embedded in the microscope bath filled with 8 mL of imaging medium.

Geoghegan N.D., Evelyn C., Whitehead L.W., Pasternak M., McDonald P., Triglia T., Marapana D.S., Kempe D., Thompson J.K., Mlodzianoski M.J., Healer J., Biro M., Cowman A.F, & Rogers K.L. (2021). 4D analysis of malaria parasite invasion offers insights into erythrocyte membrane remodeling and parasitophorous vacuole formation. Nature Communications, 12, 3620.

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Apoptosis Pathway Inhibition Evaluation

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ABT-737, z-VAD-FMK, z-DEVD-FMK, and z-LEHD-FMK were obtained from BioVision. Trolox and CTSI (cathepsin inhibitor) were obtained from Santa Cruz. E-64, chloroquine, and N-acetylcysteine were obtained from Sigma-Aldrich.
The antibodies used in this study are as follows: caspase 9 (Cat. #9508; Cell Signaling Technology), cathepsin B (Cat. #ab58802; Abcam), Bcl-2 (Cat. #ab692; Abcam), and Bcl-xL (Cat. #ab77571; Abcam).

Yin P., Jia J., Li J., Song Y., Zhang Y, & Chen F. (2016). ABT-737, a Bcl-2 Selective Inhibitor, and Chloroquine Synergistically Kill Renal Cancer Cells. Oncology Research, 24(1), 65-72.

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KGN Cell Culture and Stimulation

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Procedures have recently been described [2 (link)]. The patented KGN cell line was obtained from RIKEN BioResource Center [4 (link)] with permission by T. Yanase. KGN cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM)/Ham’s F12 medium (Life Technologies, Paisley, UK) supplemented with penicillin (100 U/mL), streptomycin (100 μg/mL) (Biochrom, Berlin, Germany) and 10% fetal calf serum (FCS) (Capricorn Scientific, Ebsdorfergrund, Germany) at 37 °C and with 5% CO₂. For stimulation experiments, 20 µM Trolox (Santa Cruz Biotechnology, Dallas, TX, USA) or 100 µM/1 mM H₂O₂ (Sigma-Aldrich, St. Louis, MO, USA) was diluted in DMEM/Ham’s F12 medium (Life Technologies; colorless medium without phenol red was used for live cell fluorescence imaging to reduce background autofluorescence).

Hack C.T., Buck T., Bagnjuk K., Eubler K., Kunz L., Mayr D, & Mayerhofer A. (2019). A Role for H2O2 and TRPM2 in the Induction of Cell Death: Studies in KGN Cells. Antioxidants, 8(11), 518.

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Optimized Buffers for Origami and Cell Imaging

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For imaging origami structures, a Tris-based buffer (5 mM Tris, 10 mM MgCl₂, 1 mM EDTA, 0.05% Tween 20, 20 mM Na₂SO₃ and 1 mM Trolox, pH 7.3-7.5) was prepared. For imaging fixed cell samples, a high ionic strength PBS-based buffer (1× PBS, 500 mM NaCl, 20 mM Na₂SO₃ and 1 mM Trolox, pH 7.3-7.5) was used. Trolox (Santa Cruz Biotechnology, sc-200810) aliquots were stored at −20°C at a concentration of 50 mM in DMSO and thawed prior to the experiment. Imager probes were stored at −20°C at a concentration of 100 μM in nuclease-free H₂O and serially diluted into one of the imaging buffers as necessary.

Chung K.K., Zhang Z., Kidd P., Zhang Y., Williams N.D., Rollins B., Yang Y., Lin C., Baddeley D, & Bewersdorf J. (2022). Fluorogenic DNA-PAINT for faster, low-background super-resolution imaging. Nature methods, 19(5), 554-559.

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Amplex Red H2O2/Peroxidase Assay Protocol

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The Amplex Red Hydrogen Peroxide/Peroxidase Assay Kit including 3% H₂O₂ for the establishment of standard curve (Cat # A-22188), Invitrogen 96-well microplates for fluorescence-based assays (Cat # M33089), dihydroethidium (DHE) (Cat # D1168), Hoechst 33342 (Cat # H3570), and MitoSOX Red mitochondrial superoxide indicator (Cat # M36008) were purchased from Invitrogen. Trolox (Cat # SC-200810) was obtained from Santa Cruz Biotechnology. Lipopolysaccharides (LPS) from Escherichia coli O111:B4 (Cat # L4391), Rosmarinic acid (RA) (Cat # 536954-5G), N-acetyl cysteine (NAC) (Cat # A9165), Vitamin C (Cat # A4403), and sodium pyruvate (Cat # P5280) were purchased from Sigma-Aldrich. WST-1 (Cat # 11644807001) for cell viability assays was obtained from Roche Applied Science. Rat anti-mouse CD3 pacific blue (Cat #100334), CD4 FITC (Cat # 100406), CD8 Percp (Cat # 100732), CD44 APC (Cat # 103012), CD62L PE/Cy7 (Cat # 104418), IFN-γ PE (Cat # 505807), IL-17 PE/Cy7 (Cat # 506921), CD103 PE (Cat # 121406), CD86 Pacific blue (Cat # 105021), I-A/I-E FITC (Cat # 107605), CD40 APC (Cat # 124611), anti-mouse CD16/32 (Cat # 156603), hamster anti-mouse CD11c PerCP (Cat # 117326), hamster anti-mouse CD80 PE/Cy7 (Cat # 104734) and mouse anti-mouse H-2Kd/H-2Dd PE (Cat # 114708) were purchased from BioLegend.

Guo S., Burcus N.I., Scott M., Jing Y, & Semenov I. (2021). The role of reactive oxygen species in the immunity induced by nano-pulse stimulation. Scientific Reports, 11, 23745.

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Thiamine Deficiency Assessment Protocol

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Amprolium, Trolox, and 2,3,5‐triphenyl‐tetrazolium chloride (TTC) were obtained from Santa Cruz Biotechnology (CA, USA). AIN‐93M (standard) and AIN‐93TD (thiamine deficient) chow were purchased from PRAG Soluções Biociências (SP, Brazil). Dimethyl sulfoxide (DMSO) was purchased from Amresco (OH, USA). HEPES was obtained from Uniscience (SP, Brazil). All other reagents were of the highest analytical grade.

Moraes J.O., Rodrigues S.D., Pereira L.M., Medeiros R.D., de Cordova C.A, & de Cordova F.M. (2018). Amprolium exposure alters mice behavior and metabolism in vivo. Animal Models and Experimental Medicine, 1(4), 272-281.

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Comprehensive Characterization of Phytochemicals

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HPLC-grade acetonitrile (CH₃CN), Folin-Ciocalteu phenol reagent, aluminum trichloride, hydrochloric acid, and sodium carbonate were purchased from Merck (Darmstadt, Germany). Methanol (MeOH), ethanol, 2-propanol, potassium acetate, orthophosphoric acid, sodium dihydrogen phosphate, potassium hydroxide, and hexane were obtained from J.T. Baker Chemical Co. (Phillipsburg, NJ, USA). Tannic acid, quercetin, gallic acid, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were provided from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Trolox was supplied by Santa Cruz Biotechnology. L-Ascorbic acid and amino acid standard 17 types were procured from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA).

Coloma A., Flores-Mamani E., Quille-Calizaya G., Zaira-Churata A., Apaza-Ticona J., Calsina-Ponce W.C., Huata-Panca P., Inquilla-Mamani J, & Huanca-Rojas F. (2022). Characterization of Nutritional and Bioactive Compound in Three Genotypes of Mashua (Tropaeolum tuberosum Ruiz and Pavón) from Different Agroecological Areas in Puno. International Journal of Food Science, 2022, 7550987.

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