All cell culture media components were purchased from Invitrogen (Paisley, UK). Dnmt1 KO and Dnmt3a/3b double KO cells with matching WT were kind gifts from Dr. Masaki Okano (RIKEN Center for Developmental Biology, Kobe, Japan). ESCs were maintained on Nunc plates (Davidson & Hardy, Belfast, UK) treated with 0.1% gelatin (Sigma-Aldrich, Dorset, UK) and cultured in Knockout DMEM plus 15% knockout serum replacement, 1% ESC-qualified Foetal Bovine Serum, 1× NEAA, 2 mM l -glutamine, 0.1 mM β-mercaptoethanol (Sigma-Aldrich, Dorset, UK) and 1000U/ml LIF (Merck Millipore, Hertfordshire, UK).
Cell culture media components
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Cell culture media components are essential ingredients used in the preparation of cell culture media. These components provide the necessary nutrients, growth factors, and other essential elements required for the growth, maintenance, and proliferation of cells in a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Foetal bovine serum (fbs), by Merck Group (1 mentions)
Knockout dmem, by Merck Group (1 mentions)
Gelatin, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Chroma spin 10 columns, by Takara Bio (1 mentions)
Polyvinylidene difluoride pvdf membrane, by Bio-Rad (1 mentions)
Imr 32 cells, by American Type Culture Collection (1 mentions)
Neutral red dye, by Merck Group (1 mentions)
Resazurin, by Merck Group (1 mentions)
Low melting point agarose, by Merck Group (1 mentions)
Tris hydrochloride tris hcl, by Merck Group (1 mentions)
Sybergold, by Thermo Fisher Scientific (1 mentions)
Absolute ethanol c2h6o, by Merck Group (1 mentions)
Methyl methanesulfonate mms, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Potassium hydroxide, by Merck Group (1 mentions)
3 protocols using cell culture media components
1
Culturing Dnmt-Deficient Embryonic Stem Cells
2
Modulation of STAT1/STAT3 Signaling in IMR-32 Cells
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IMR-32 cells were obtained from the American Type Culture Collection (Rockville, MA). Cell culture media components were obtained from Invitrogen Life Technologies (Carlbad, CA). Antibodies for STAT1, STAT3, β-actin, α-tubulin, β-tubulin and heterogeneous nuclear ribonucleoprotein A1 (hnRNP), and the oligonucleotides containing the consensus sequence for STAT1 and STAT3 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). The antibody for 4-hydroxynonenal was purchased from Abcam (Cambridge, MA). Antibodies for phosphotyrosine-701 STAT1 (pY701-STAT1), phosphoserine-727 STAT1 (pS727-STAT1), phosphotyrosine-705 STAT3 (pY705-STAT3), and phosphoserine-727 STAT3 (pS727-STAT3) were purchased from Cell Signaling Technologies (Danvers, MA). The reagents for EMSA assay were obtained from Promega (Madison, WI). Polyvinylidene difluoride (PVDF) membranes were obtained from Bio-Rad (Hercules, CA, USA). Chroma Spin-10 columns were obtained from Clontech (Mountain View, CA). Vinblastine (Vb), colchicine (Col), cytochalasin D (Cyt D), α-lipoic acid (LA) and all other reagents were of the highest quality available and were purchased from Sigma (St. Louis, MO).
3
Cytotoxicity Assay Protocol
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3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), neutral red dye, resazurin, Triton X-100, 2-mercaptoethanol, low melting point (LMP) agarose, Tris hydrochloride (Tris-HCl) and methyl methanesulfonate (MMS) were purchased from Sigma-Aldrich Co. Dimethyl sulfoxide (DMSO), glacial acetic acid (CH 3 COOH), absolute ethanol (C 2 H 6 O), sodium hydroxide (NaOH), sodium chloride (NaCl) and potassium hydroxide (KOH) were bought from Merck KGaA. Cell culture media components and SyberGold® were all Invitrogen™ and purchased from Thermo Fisher Scientific Inc. Lactate dehydrogenase (LDH) protein from rabbit muscle and Tris base were purchased from Calbiochem, while potassium chloride (KCl) and disodium salt dihydrate (Na 2 EDTA) were purchased from Prolab. Finally, high purity
and normal melting point (NMP) agarose were supplied from VWR chemicals, Lonza Group AG and Bioline, respectively.
and normal melting point (NMP) agarose were supplied from VWR chemicals, Lonza Group AG and Bioline, respectively.
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