Every time a new batch of EBRTcPTbx6 cells was thawed, 1.5 μg/mL of puromycin was added for one week before propagation. EBRTcPTbx6 cells were maintained with Glasgow minimum essential medium (GMEM, Sigma–Aldrich), 10% FBS (MP Biomedicals), 0.1 mM NEAA (Wako), 1 mM sodium pyruvate (Sigma–Aldrich), 0.1 mM 2-mercaptoethanol (Sigma–Aldrich), 10 ng/mL doxycycline (Wako), 100 U/mL penicillin (Meiji Seika Pharma) and 100 μg/mL streptomycin (Meiji Seika Pharma), supplemented with 1000 U/mL mouse LIF (ORF genetics), 3 μM CHIR-99021 (FOCUS Biomolecules), and 0.4 μM PD-0325901 (Adooq Bio Science). 1 × 105 EBRTcPTbx6 cells were inoculated to a 60 mm cell culture dish (TrueLine, Nippon Genetics) that had been plated with MMC (Wako)-treated SNL 76/7 feeder cells (obtained from Riken Bioresource Center) on the previous day. The cells were passaged every other day, and subjected to isolation from feeder cells using differential adhesive properties to a gelatin-coated cell culture dish before performing differentiation assays as described below.
Pd 0325901
Manufactured by Adooq
Sourced in United States
PD-0325901 is a laboratory instrument used for the analysis and examination of biological samples. It is designed to perform specific functions related to the study and evaluation of such samples. The core function of this product is to facilitate the research and investigation of various biological materials.
Automatically generated - may contain errors
Lab products found in correlation
Non essential amino acids (neaa), by Fujifilm (2 mentions)
Doxycycline, by Fujifilm (1 mentions)
Chir 99021, by Focus Biomolecules (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Sodium pyruvate, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by MP Biomedicals (1 mentions)
Trueline, by Nippon Genetics (1 mentions)
Penicillin, by Meiji Seika Pharma (1 mentions)
Streptomycin, by Meiji Seika Pharma (1 mentions)
Leukemia inhibitory factor, by Fujifilm (1 mentions)
1 thioglycerol, by Merck Group (1 mentions)
Stemsure d mem, by Fujifilm (1 mentions)
Chir99021, by Cayman Chemical (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Nacalai Tesque (1 mentions)
Trypsin, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
2 protocols using pd 0325901
1
Expansion of Murine Embryonic Stem Cells
2
Culturing Mouse Induced Pluripotent Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Mouse iPS cells (iPS‐Mef‐Ng‐492B‐4) [26 (link)] from RIKEN BRC cell bank were grown on feeder layers of mitomycin C‐treated mouse embryonic fibroblasts (MEF) derived from E14.5 CD1 mice on gelatin‐coated dishes in stem sure DMEM (Wako, Osaka, Japan) supplemented with 15% stem sure serum replacement (SSR; Wako), 1% non‐essential amino acids (NEAA; Wako), 1% Glutamax (Life Technologies, Carlsbad, CA, USA), 0.5 mm 1‐thioglycerol (Sigma‐Aldrich, St. Louis, MO, USA), 1000 units·mL−1 leukemia inhibitory factor (Wako), 3 μm CHIR99021 (Cayman Chemical, Ann Arbor, MI, USA), 1 μm PD0325901 (AdooQ Bioscience, Irvine, CA, USA) and 1% penicillin/streptomycin (Nacalai Tesque, Kyoto, Japan) at 37 °C in a humidified 5% CO2. For routine passaging, mouse iPS colonies were dissociated enzymatically with 0.25% trypsin and 0.1% EDTA (Sigma‐Aldrich) in PBS.
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