CoCl2·6H2O was purchased from Sigma Chemical Co (USA). Isoproterenol Yi Feixue Bio, Nanjing, China. Hydroxytyrosol (98%) was purchased from Aladdin Biological Reagent Company (Shanghai, China). OLE (25%) was provided by Sinolife United Biotech Company. H9c2 cells were obtained from the Cell Bank of Shanghai Institute of Biochemistry and Cell Biology. Dulbecco's Modified Eagle's Medium (DMEM) and FBS were purchased from ScienCell Research Laboratories (CA, USA). Primary antibodies against GRP78, CHOP and β-actin were purchased from Cell Signaling Technology (Danvers, MA, USA). Goat anti-rabbit IgG and goat anti-mouse IgG antibodies were purchased from ZSGB-BIO (Beijing, China). Hochest 33258 was obtained from the Beyotime Institute of Biotechnology (Shanghai, China). Primers used for quantitative RT-PCR were provided by Invitrogen (Carlsbad, Calif, USA). GRP78:(Forward-5'-TCAGCCCACCGTAACAATCAAGG-3'; Reverse-5'-CTTCCTCAGCAAACTTCTCGGCG-3'). CHOP: (Forward-5'-GCACCTCCCAAAGCCCTCGC-3'; Reverse-5'-CCGTTTCCTAGTTCTTCCTT-3')
Dulbecco s modified eagle s medium dmem
Manufactured by ScienCell
Sourced in United States
Dulbecco's Modified Eagle's Medium (DMEM) is a widely used cell culture medium formulation designed to support the growth and maintenance of a variety of cell types. It provides essential nutrients, vitamins, and salts required for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by ScienCell (2 mentions)
Goat anti rabbit igg, by ZSGB-BIO (1 mentions)
Cocl2 6h2o, by Merck Group (1 mentions)
Hochest 33258, by Beyotime (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Goat anti mouse igg, by ZSGB-BIO (1 mentions)
Primovert, by Zeiss (1 mentions)
Penicillin streptomycin, by ScienCell (1 mentions)
L glutamine, by ScienCell (1 mentions)
Fetal bovine serum (fbs), by Beyotime (1 mentions)
Ht22 cells, by Procell (1 mentions)
Penicillin streptomycin mixture, by BasalMedia (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Foetal bovine serum (fbs), by Merck Group (1 mentions)
Stat1, by Thermo Fisher Scientific (1 mentions)
Secondary antibody conjugated to horseradish peroxidase, by Thermo Fisher Scientific (1 mentions)
4 protocols using dulbecco s modified eagle s medium dmem
1
Endoplasmic Reticulum Stress Modulation
2
Isolation and Expansion of Human Fibroblasts
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Primary cultures of human dermal and pulmonary fibroblasts (obtained from ScienCell, Carlsbad, CA, US) were grown under standard conditions (37 °C, 5% CO2) in Dulbecco’s modified Eagles medium (DMEM), supplemented with 10% fetal bovine serum, 1% L-glutamine, and 1% penicillin/streptomycin. All products for cell cultures were from Biological Industries, Beit Haemek, Israel. The cultures were inspected every day under inverted phase-contrast microscope (Primo Vert, Zeiss) and cells were passaged 1:2 upon 75–80% confluence. The number and concentration of viable cells were calculated using Trypan blue exclusion assay. Replicative CS was achieved by serial passaging. The cells were defined as pre-senescent or senescent, based on a dramatic inhibition of cell proliferation or cell growth arrest, respectively; typical CS morphology, and expression of the CS marker SA-β-gal. The SA-β-gal assay was carried out using the Sigma Aldrich SA-β-galactosidase detection kit (GALS), according to the manufacturer’s protocol, with subsequent visualization using the Primo Vert microscope.
3
Culturing Mouse Hippocampal and Astrocyte Cells
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Mouse hippocampal neuron HT22 cells were purchased from Procell Life Science & Technology Co., Ltd. (cat. no. CL-0595). MA cells were purchased from Jennio Biotech Co., Ltd. (cat. no. JNO-M0088), which were immortalized from mouse primary astrocytes (cat. no. 1800-57) provided by ScienCell Research Laboratories, Inc. HT22 cells and MA cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM; cat. no. D211113; Shanghai BasalMedia Technologies Co., Ltd.) supplemented with 1% penicillin/streptomycin mixture (cat. no. C0222; Beyotime Institute of Biotechnology) and 10% fetal bovine serum (FBS; cat. no. 11011-8611; Zhejiang Tianhang Biotechnology Co., Ltd.). The cells were routinely incubated in an incubator containing 5% CO2 at a temperature of 37˚C. When cells reached an 80-90% confluency, they were subcultured at a 1:2 ratio using trypsin (cat. no. J121002; Shanghai BasalMedia Technologies Co., Ltd.).
4
Aflatoxin B1 Cytotoxicity Assay
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The AFB1 used in the study was purchased from Sigma-Aldrich, USA (cat no A6636) and dissolved in dimethyl sulfoxide (DMSO) to a stock concentration of 3200 μM. The AFB1 stock solution was divided into aliquots, wrapped in aluminium foil and stored frozen at −20 °C until used. The AFB1 stock solution was diluted to the desired concentration in normal growth medium when necessary. The foetal bovine serum (FBS) was purchased from Sigma-Aldrich, USA. Dulbecco’s Modified Eagles Medium (DMEM) (high glucose, L-glutamine, sodium pyruvate and 25 mM HEPES) was purchased from Science Cell. Minimum essential medium (MEM) non-essential amino acids was purchased from Sigma Aldrich, USA while penicillin-streptomycin was purchased from Gibco by Invitrogen, UK. Lipofectamine 2000 was purchased from Gibco by Life Technologies, UK (cat no 11668-019). The human recombinant interferon-alpha 2 (rIFN-α2) was purchased from PBL interferon source (cat no 11115-1). The stock solution of the human recombinant interferon-alpha 2 was diluted to working concentration using phosphate buffered saline containing 0.1% bovine serum albumen as a diluent. The STAT1 (cat no PA5-34504) and GAPDH (cat no QE 212271) primary antibodies were purchased from Thermo Scientific, USA. The secondary antibody conjugated to horse-radish peroxidase (cat no 31430) was purchased from Thermo Scientific, USA.
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