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F6178

Manufactured by Merck Group
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The F6178 is a laboratory equipment product manufactured by Merck Group. It is designed for general laboratory use, but the specific core function of the equipment is not available in the information provided. A detailed and unbiased description cannot be provided while maintaining the required conciseness.

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Lab products found in correlation

D6429, by Merck Group (3 mentions) Penicillin streptomycin, by Merck Group (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) 6230b accurate mass tof lc ms system, by Agilent Technologies (1 mentions) P4333, by Merck Group (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Dmem hg, by Merck Group (1 mentions) H1138, by Merck Group (1 mentions) D5796, by Merck Group (1 mentions) Mt10013cv, by Merck Group (1 mentions) Penicillin, by Corning (1 mentions) Streptomycin, by Corning (1 mentions) Pen strep, by Corning (1 mentions) Human cystatin c duoset catalog dy1196, by R&D Systems (1 mentions) Iscove s modified dulbecco s medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)

16 protocols using f6178

1

Propagation of Rickettsia in Vero Cells

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Vero cells (ATCC CCL-81) were used as the host cells for Rickettsia spp. infection as described previously.1 The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Sigma D6429) with 10% fetal bovine serum (FBS, Sigma F6178) and grown at 37 °C with 5% CO2. The cells were passaged every 2–3 days and harvested or passed at between 80% and 95% confluency following previously established protocols.
Lemenze A., Mittal N., Perryman A.L., Daher S.S., Ekins S., Occi J., Ahn Y.M., Wang X., Russo R., Patel J.S., Daugherty R.M., Wood D.O., Connell N, & Freundlich J.S. (2022). Rickettsia Aglow: A Fluorescence Assay and Machine Learning Model to Identify Inhibitors of Intracellular Infection. ACS infectious diseases, 8(7), 1280-1290.
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2

Quantitative Metabolomic Analysis of Vero Cells

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Vero cells were suspended in Dulbecco’s modified Eagle’s medium (Sigma-Aldrich D6429) supplemented with 10% fetal bovine serum (Sigma-Aldrich F6178) to a concentration of 0.5–1.0 × 106 cells/mL with the cell viability >90%. The assay was initiated by treating the Vero cells with a compound at 5.0 or 10 μM in the six-well plate in triplicate. DMSO-treated culture was used as a control. Each well containing Vero cells was washed with ice-cold PBS after discarding the media. Then, the cells were collected by scraping and resuspended with prechilled 1 mL of solvent (2:2:1 CH3CN/MeOH/H2O). The resuspended pellet was lysed by four freeze–thaw cycles at −80 °C. After lysis, the samples were centrifuged at 14,000 rpm, 5 min, 4 °C, and the supernatant was filtered through a 0.22 μm centrifugal filter unit. An aliquot of filtrate was injected onto a Chromolith SpeedRod column, with a gradient of acetonitrile in water (10–100%) acidified with 0.1% formic acid, interfaced with an Agilent 6230B Accurate Mass TOF LC/MS system. The mass resolution consisted of m/z = 100–20,000 with sub-ppm accuracy within the calibrated mass range. Concentrations were calculated using the standard curve for the compound with verapamil as an internal standard.
Lemenze A., Mittal N., Perryman A.L., Daher S.S., Ekins S., Occi J., Ahn Y.M., Wang X., Russo R., Patel J.S., Daugherty R.M., Wood D.O., Connell N, & Freundlich J.S. (2022). Rickettsia Aglow: A Fluorescence Assay and Machine Learning Model to Identify Inhibitors of Intracellular Infection. ACS infectious diseases, 8(7), 1280-1290.
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3

Standardized Fibroblast Culture for Seahorse Analysis

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PD patients’ fibroblasts were prepared isolated at Leiden University Medical Center from skin biopsies derived from the ventral side of the upper leg and cultured under highly standardized conditions at 37°C and 5% Carbon dioxide (CO2) up to a maximum of 10 passages. The number of passages was kept consistent within groups. Fibroblasts were maintained in Dulbecco's Modified Eagle Medium (DMEM) 10% Fetal Bovine Serum (FBS) and 1% penicillin‐streptomycin (P4333, Sigma‐Aldrich) until reaching confluency. Prior to Seahorse analysis, fibroblasts were rinsed with Phosphate‐Buffered Saline (PBS) and cultured with glucose (glucose 10 mM, 10% FBS [F6178, Sigma‐Aldrich], 2 mM glutamine, 5 mM Hepes, and 1% penicillin‐streptomycin (P4333, Sigma‐Aldrich, St. Louis, MI, USA) or galactose (galactose 10 mM, 10% FBS, 2 mM glutamine, 5 mM Hepes, and 1% penicillin‐streptomycin) medium for 3 days. Fibroblasts from sex‐matched controls of comparable age were either obtained from the Coriell biorepository (identification codes AG04659, AG05266, AG06283, AG07936, AG08152, AG08268, AG08269, AG08543, AG09162, AG09271, AG09879, AG12428, AG12951, AG13077, AG13348), Coriell Institute for Medical Research (Camden, New Jersey), or from the biorepository available in the Department of Molecular Genetics of the Erasmus Medical Center (MC), Rotterdam, The Netherlands.
Milanese C., Payán‐Gómez C., Galvani M., Molano González N., Tresini M., Nait Abdellah S., van Roon‐Mom W.M., Figini S., Marinus J., van Hilten J.J, & Mastroberardino P.G. (2019). Peripheral mitochondrial function correlates with clinical severity in idiopathic Parkinson's disease. Movement Disorders, 34(8), 1192-1202.
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4

Cas9 expression in cell lines

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L929 (CCL1-1, ATCC), HT-29 (HTB-38), and COLO 205 (CCL-222) cells were modified to express human codon-optimized Cas9 introduced using the blasticidin selectable pLENTI6.3 Cas955 (link). L929.pLENTI6.3-Cas9 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS; F6178, Sigma), 2 mM Glutamax (35050061, ThermoFisher), 100 IU penicillin, and 100 μg/ml streptomycin (15140122, Thermofisher), 1% non-essential amino acids (11140050, ThermoFisher), 10 mM HEPES pH 7.2 and 6 μg/ml blasticidin (A1113903, ThermoFisher). HT-29.pLENTI6.3-Cas9 and COLO 205.pLENTI6.3-Cas9 cells were cultured in RPMI1640 supplemented with 10% FBS and 10 μg/ml blasticidin. HEK-293T cells were cultured in DMEM supplemented with 10% FBS, 2 mM Glutamax, and 1% non-essential amino acids. Human iPSC-derived neural stem cells (MTI-Global Stem, GSC-4311) were maintained in NSC maintenance medium (DMEM/F12/Neurobasal, 1× GS22, 20 ηg/ml bFGF, 20 ηg/ml EGF, 20 ηg/ml BDNF, 0.11 mM β-mercaptoethanol, 0.5 mM Glutamax) and differentiated in NSC differentiation medium (DMEM/F12/Neurobasal, 1× GS21, 1× N2, 5 μg/ml cholesterol, 1 mM creatine, 100 μM ascorbic acid, 0.5 mM cAMP, 20 ηg/ml GDNF, 20 ηg/ml BDNF, 1 μg/ml laminin, 0.5 mM Glutamax). iPSC neural conversion was monitored by microscopic inspection of cells.
Callow M.G., Watanabe C., Wickliffe K.E., Bainer R., Kummerfield S., Weng J., Cuellar T., Janakiraman V., Chen H., Chih B., Liang Y., Haley B., Newton K, & Costa M.R. (2018). CRISPR whole-genome screening identifies new necroptosis regulators and RIPK1 alternative splicing. Cell Death & Disease, 9(3), 261.
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5

C2C12 Myoblast Cell Culture

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All medium was based on Dulbecco’s Modified Eagle’s Medium-high glucose (DMEM-HG cat no. D5796, Sigma-Aldrich). Components added to the medium were sterile filtered with syringe filters with a 0.45 µm pore size (GE Healthcare, Whatman). The myoblast cell line C2C12 (DSMZ, ACC 565) was maintained at 37°C in 5% CO2 at 95% humidity in growth medium (GM) containing DMEM-HG (Sigma-Aldrich), 10% fetal bovine serum (F6178, Sigma-Aldrich), and 1% penicillin/streptomycin (15140122, Thermo Fisher Scientific). Differentiation medium (DM) was prepared from DMEM-HG with 2% horse serum (H1138, Sigma-Aldrich) and 1% penicillin/streptomycin.
Mosqueira M., Scheid L.M., Kiemel D., Richardt T., Rheinberger M., Ollech D., Lutge A., Heißenberg T., Pfitzer L., Engelskircher L., Yildiz U, & Porth I. (2022). nNOS-derived NO modulates force production and iNO-derived NO the excitability in C2C12-derived 3D tissue engineering skeletal muscle via different NO signaling pathways. Frontiers in Physiology, 13, 946682.
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6

Isolation of Stromal Vascular Fraction from Adipose Tissue

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Consent forms for adipose tissue donation were prepared in accordance with the University of Johannesburg’s’ institutional guidelines. Ethical approval (REC-01–186-2016) was obtained from the Research Ethics Committee of the Faculty of Health Sciences, University of Johannesburg in accordance with the Human Tissue Act 65, 1983). Tissue was obtained from a donor undergoing abdominoplasty in Linksfield hospital, Johannesburg, South Africa and the isolation was performed using the collagenase digestion method [17 (link)]. Briefly, donated tissue was subjected to enzymatic digestion with collagenase solution, followed by high-speed centrifugation and filtration of the infranatant through a 40 μm filter (BD Biosciences, 352,340) to obtain the stromal vascular fraction (SVF) containing high numbers of blood cells with traces of endothelial cells, fibroblasts, pericytes and stem cells. Cell pellet containing the SVF was re-suspended in Erythrocyte Lysis Buffer to remove blood cells and the solution was then spun at 0.5 ×g for 5 min. The residue was re-suspended in a complete culture medium consisting of Dulbecco’s Modified Eagle Medium (DMEM, Sigma, D5796) supplemented with 10% foetal bovine serum (FBS, Sigma, F6178), 0.1% v/v penicillin/streptomycin, and 0.5% v/v fungizone for incubation at 37 °C in a humidified chamber containing 5% CO2.
George S., Hamblin M.R, & Abrahamse H. (2018). Effect of red light and near infrared laser on the generation of reactive oxygen species in primary dermal fibroblasts. Journal of photochemistry and photobiology. B, Biology, 188, 60-68.
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7

Epicardial-Derived Cell Line Control

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An epicardial-derived cell line(Russell et al., 2011 (link)) was used as a control in ICC experiments. The cell line was grown in growth media (1 volume Media 199 + 3 volumes DMEM + 10% FBS, Sigma F6178) until confluency before being harvested for ICC.
Duan J., Li B., Bhakta M., Xie S., Zhou P., Munshi N.V, & Hon G.C. (2019). Rational Reprogramming of Cellular States by Combinatorial Perturbation. Cell reports, 27(12), 3486-3499.e6.
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8

Cell Culture of HEK 293T and MCF7 Cells

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HEK 293T and MCF7 cells were utilized in the described experiments (American Type Culture Collection, Manassas, VA). HEK 293T and MCF7 cells were cultured in DMEM (Corning, MT10013CV, Manassas, VA) with 10% FBS (Sigma, F6178, St. Louis, MO) and 1% pen/strep at 5000 I.U/ml penicillin and 5000 μg/ml streptomycin (Corning, 30-001-Cl, Manassas, VA). Between passages, cells were trypsinized with 0.25% Trypsin with 2.21 mM EDTA (Corning, 25-053-Cl, Manassas, VA). All cells were maintained at 37°C in a humidified atmosphere of 5% CO2.
Umstead M., Xiong J., Qi Q., Du Y, & Fu H. (2017). Aurora kinase A interacts with H-Ras and potentiates Ras-MAPK signaling. Oncotarget, 8(17), 28359-28372.
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9

Cell Line Maintenance and Culture

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HEK293 cells, Flag-MLL-AFF1 and Flag-MLL-AF9 stable cell lines (Lin et al., 2010 (link)) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS, catalog No. F6178, Sigma). The 293C6 stable cell line with the overexpression of IL1R1 and IL1RAP in HEK293 cells was a gift from George Stark’s Lab (Cleveland Clinic) and maintained in DMEM medium with 5% FBS. MONO-MAC-6 (MM6, ACC-252), RL (CRL-2261), U-937 (CRL-1593.2), RS4;11 (CRL-1873), SU-DHL-5 (CRL-2958), SU-DHL-6 (CRL-2959), MOLM13 (ACC-554), OCI-LY1 (ACC-722), THP-1 (TIB-202), ML-2 (ACC-15), NB-4 (ACC-207) and REH (ACC-22) leukemia cells were maintained in RPMI-1640 medium supplemented with 10% FBS. MV4–11 (CRL-9591) and SEM (ACC-546) leukemia cells were maintained in Iscove’s Modified Dulbecco’s Medium (IMDM) with 10% FBS. The Mll wild-type Mll+/+ and knockout Mll−/− mouse embryonic fibroblasts (MEF) cell lines were provided by Dr. Jay Hess (University of Michigan Medical School) and cultured in DMEM with 10% FBS.
Liang K., Volk1 A.G., Haug J.S., Marshall S.A., Woodfin A.R., Bartom E.T., Gilmore J.M., Florens L., Washburn M.P., Sullivan K.D., Espinosa J.M., Cannova J., Zhang J., Smith E.R., Crispino J.D, & Shilatifard A. (2017). Therapeutic targeting of MLL degradation pathways in MLL-rearranged leukemia. Cell, 168(1-2), 59-72.e13.
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10

Estimating GFR from Cystatin C

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GFR is typically estimated from plasma creatinine and/or cystatin C using population-based equations; we calculated eGFR from plasma cystatin C using the 2012 CKD-EPI Cystatin C equation (Inker et al., 2012 (link)). Plasma cystatin C was measured by ELISA according to the manufacturer’s protocol (R&D Systems Human Cystatin C Duoset Catalog# DY1196). We used a 6 point standard curve with a high standard of 3000 pg/ml. Samples were diluted 1:2000 in PBS with 10% fetal bovine serum (Sigma Aldrich F6178). Recovery of the IFCC certified reference material for serum cystatin C (ERM-DA 471/IFCC) was 104%. The intra- and inter-assay CVs were 7% and 10%, respectively.
Peters B.A., Hall M.N., Liu X., Slavkovich V., Ilievski V., Alam S., Siddique A.B., Islam T., Graziano J.H, & Gamble M.V. (2015). Renal function is associated with indicators of arsenic methylation capacity in Bangladeshi adults. Environmental research, 143(0 0), 123-130.
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