Cell culture supplements were purchased from Invitrogen (Carlsbad, CA, USA). Antibodies against IL-6 (SC-28343), p-p65 (Ser536; SC-101752), p-65 (SC-8008), p-85 (SC-1637), p-AKT (Thr308; SC-16646-R), AKT (SC-5298), p-IKKα/β (Ser180/Ser181; SC-23470-R), IKKα/β(SC-7607), p-IκBα (Ser32; SC-8404), IκBα (SC-203), PCNA(SC-36), and β-actin (SC-58673), and C6 ceramide (an ERK activator), were all purchased from Santa Cruz (Santa Cruz, CA, USA). Anti-TNF-α (a11534) was obtained from Abclonal (Woburn, MA, USA). Antibody against p-p85 (Tyr458/Tyr199; 4228S) was purchased from Cell Signaling (Danvers, MA, USA). AKT inhibitor were supplied by Calbiochem (San Diego, CA, USA). Ly294002, TPCK and PDTC were bought from Enzo Life Sciences International (Plymouth Meeting, PA, USA) and Cell culture supplements were purchased from Invitrogen (Carlsbad, CA, USA). Dual-Luciferase® Reporter Assay System was bought from Promega (Madison, WI, USA). All other chemicals not described above were supplied by Sigma-Aldrich (St Louis, MO, USA).
Cell culture supplements
Manufactured by Thermo Fisher Scientific
Sourced in United States
Cell culture supplements are a class of laboratory products designed to support the growth and maintenance of cell cultures in vitro. These supplements provide essential nutrients, growth factors, and other components necessary for the optimal health and proliferation of cultured cells.
Automatically generated - may contain errors
Lab products found in correlation
Taqman one step pcr master mix, by Thermo Fisher Scientific (6 mentions)
Dual luciferase reporter assay system, by Promega (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Qpcr primers and probes, by Thermo Fisher Scientific (3 mentions)
On targetplus sirna, by Horizon Discovery (3 mentions)
Annexin 5 fitc propidium iodide apoptosis detection kit, by Thermo Fisher Scientific (2 mentions)
β actin sc 58673, by Santa Cruz Biotechnology (2 mentions)
Plasmid purification kit, by Thermo Fisher Scientific (2 mentions)
Cell culture media, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Disposable tissue culture supplies, by Thermo Fisher Scientific (2 mentions)
Epoxomicin, by Apexbio (1 mentions)
Tnfr2, by Cell Signaling Technology (1 mentions)
Tnf α, by Santa Cruz Biotechnology (1 mentions)
P p38 mapk, by Cell Signaling Technology (1 mentions)
Sirt3, by Cell Signaling Technology (1 mentions)
α tubulin, by Cell Signaling Technology (1 mentions)
Gkt137831, by MedChemExpress (1 mentions)
Glx351322, by MedChemExpress (1 mentions)
Mitosox, by MedChemExpress (1 mentions)
23 protocols using cell culture supplements
1
Investigating Inflammatory Signaling Pathways
2
Evaluating Apoptosis-Inducing Pathways
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Without specific indication, the reagents purchased from Sigma‐Aldrich Inc. were used in the present study, and cell culture supplements were from GIBCO/Life Technologies Inc. Vincristine (VCR), SZLP1‐41 (Skp2 inhibitor), E64D (lysosomal protease inhibitor) and epoxomicin (proteasome inhibitor) were the products of Apexbio Technology LLC. GKT137831 and GLX351322 were purchased from MedChem Express; MitoSOX, annexin V‐FITC/propidium iodide (PI) apoptosis detection kit, tetramethylrhodamine (TMRM) and H2DCFDA were from Molecular Probes (Eugene, OR). Antibodies against caspase‐3 and caspase‐8, okadaic acid, Z‐IETD‐FMK and Z‐DEVD‐FMK were purchased from Calbiochem. Antibodies separately against PP2Ac, Mcl‐1, tristetraprolin (TTP), Fas, FasL, TNF‐α and FADD were purchased from Santa Cruz Biotechnology Inc.; and antibody against TNFR1 was from R&D Systems. Antibodies separately against ERK, p‐ERK, p38 MAPK, p‐p38 MAPK, JNK, p‐JNK, SIRT3, TNFR2, α‐tubulin, MID1, α4, caspase‐9, Bax, Bak and Bcl‐2 were obtained from Cell Signaling Technology. Antibodies separately against cytochrome c, Bcl‐xL and Bid were the products of BD Pharmingen Technical (San Jose, CA), and anti‐NOX4 antibody was from Novus Biologicals. Secondary antibodies conjugated with horseradish peroxidase (HRP) were the products of Pierce (Rockford, IL).
3
Apoptosis Induction and Mitochondrial ROS
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Without specific indication, the reagents obtained from Sigma-Aldrich Inc. (St. Louis, MO, USA) were used in this study, and cell culture supplements were the products of GIBCO/Life Technologies Inc. (Carlsbad, CA, USA). ABT-263 was obtained from Apexbio Technology LLC (Houston, TX, USA), and A-1210477 was the product of MedChem Express (Monmouth Junction, NJ, USA). MitoSOX Red, tetramethylrhodamine (TMRM), annexin V-FITC/propidium iodide (PI) apoptosis detection kit, and dichlorodihydrofluorescein diacetate (H2DCFDA) were obtained from Molecular Probes (Eugene, OR, USA); and Z-DEVD-FMK was from Calbiochem (San Diego, CA, USA).
4
Exosome Characterization and Isolation
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All cell culture media (DMEM, RPMI 1640) and cell culture supplements were purchased from Gibco Inc. (Billings, MT, USA), and cell culture plasticware was obtained from ThermoFisher, unless otherwise mentioned. Millipore deionized water of 18.0 Ω/cm2 was used for reagent dilution when required. MTT assay reagents (Cell Proliferation Kit I (MTT) Cat# 11465007001) were purchased from Roche. Exosome surface marker antibodies were obtained from Abcam (Cat# ab275018: Exosome Panel; CD9, CD63, CD81, TSG101, Hsp70, Calnexin). Annexin-V was purchased from BD Bioscience (Cat# 556419), and propidium iodide (PI) was obtained from Invitrogen, ThermoFisher Scientific (Waltham, MA, USA). The exosome isolation kit (total exosome isolation (cell culture), Cat# 4478359) was purchased from InvitrogenTM by Thermo Fisher Scientific.
5
Immunohistochemistry Staining Protocol
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Cell culture media (DMEM/F12, IMDM) and Growth Factor Reduced Matrigel were from Corning Inc., Corning, NY, USA. Hydrocortisone was from MilliporeSigma, Burlington, MA, USA. Cell culture supplements were from GibcoTM, Billings, MT, USA. Cell culture flasks and dishes were from Nunc, Thermo Fisher Scientific, Rochester, NY, USA; 96-well round-bottom low attachment plates were from Corning, Corning, NY, USA. Monoclonal anti-ASPH-antibody was from Santa Cruz (SC271391). DAB chromagen was from Agilent (Dako #K3468), and Hematoxylin was from Thermo Fischer, Memphis, TN, USA (Harris Modified Hematoxylin).
6
MTT Assay for Cytotoxicity in AML Cell Lines
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Human AML U937 and HL-60 cells from Bioresource Collection and Research Center (Hsinchu, Taiwan) were cultured in RPMI 1640 medium containing 10% FCS, 1% sodium pyruvate, 2 mM glutamine, and 1% penicillin/streptomycin. The cells were maintained at 37 °C in humidified air containing 5% CO2. Cell culture supplements were purchased from GIBCO/Life Technologies Inc. (Carlsbad, CA, USA). The survival rate of the cells was determined using an MTT assay. Exponentially growing cells (5 × 104) were plated in each well of a 96-well plate and treated with 0 to 1000 nM CTXs in serum-free medium for 4 h. MTT solution (1 mg/mL) was then added to each well and incubated for 4 h. After removing the supernatant from the wells, the formazan crystals produced by the reduction of MTT were dissolved by addition of 100 μL DMSO per well. The absorbance was measured at 595 nm using a plate reader.
7
Modulating PPARγ Signaling in Macrophages
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The PPARγ agonist rosiglitazone and the PPARγ antagonist GW9662 were a kind gift from Prof. Dr. von Knethen (Institute for Biochemistry I, Goethe-University Frankfurt, Germany). Conditioned medium from macrophages was supplemented and incubated with the PPARγ agonist (1 µM) or the antagonist (100 nM) for 30 min prior to stimulating NRK-52e cells with conditioned medium for 24 h. Neutralizing anti-LCN-2 and an isotype control anti-IgG were purchased from R&D (Madrid, Spain). Cell culture supplements and FCS were ordered from Gibco (Darmstadt, Germany). Primers were bought from Invitrogen (Barcelona, Spain). The siRNA against Lcn-2 and scrambled, nontargeting control siRNA were from Invitrogen. All chemicals were of the highest grade of purity and commercially available. LY294002 was ordered from Alexis Biochemical (Lörrach, Germany). A loss of cell viability was excluded for cell treatments based on trypan blue staining (Biochrom AG, Berlin, Germany).
8
Chitosan-TPP Nanoparticle Synthesis
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Water soluble 20 kDa chitosan, with a degree of deacetylation of ~70%, from Coyote Foods (Saltillo, CH, Mexico) and Pentasodium Tripolyphosphate (TPP) from Sigma–Aldrich (St. Louis, MO, USA) were weighed and resuspended in MiliQ water, at concentrations of 2 mg/mL and 0.86 mg/mL, respectively. The pH of chitosan was measured and adjusted to 5.5, while for TPP it was adjusted to 3. Cell culture media, fetal bovine serum (FBS), cell culture supplements, and plasmid purification kit were obtained from Invitrogen (Carlsbad, CAL, USA). Iron II sulfate heptahydrate and iron III chloride hexahydrate were purchased from Sigma–Aldrich (St. Louis, MO, USA), ammonium hydroxide was obtained from Productos Quimicos Monterrey (Monterrey, NL, Mexico).
9
Signaling Pathway Modulation Reagents
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Antibodies against FOXO3, AMPK, and p38 were all purchased from Santa Cruz (Santa Cruz, CA, USA). Antibodies against p-AMPK and p-p38 were purchased from Cell Signaling (Cell Signaling, UK). ON-TARGETplus siRNAs for FOXO3, AMPKα1, AMPKα2, p38 and the control were bought from Dharmacon (Lafayette, CO, USA). Ara A, Compound C and SB203580 were supplied by Calbiochem (San Diego, CA, USA). Cell culture supplements were purchased from Invitrogen (Carlsbad, CA, USA). Dual-Luciferase® Reporter Assay System was bought from Promega (Madison, WI, USA). The qPCR primers and probes, as well as the Taqman® one-step PCR Master Mix, were supplied by Applied Biosystems (Foster City, CA). All other chemicals not described above were supplied by Sigma-Aldrich (St Louis, MO, USA).
10
Cell Culture Reagents Protocol
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Unless otherwise stated, reagents were purchased from Merck Sigma Aldrich (St. Louis, MI, USA). Cell culture supplements were obtained from Invitrogen Thermo Fisher Scientific (Waltham, MA, USA).
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