2 mercaptoethanol  (Millipore)


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    Structured Review

    Millipore 2 mercaptoethanol
    2 Mercaptoethanol, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 572 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/2 mercaptoethanol/product/Millipore
    Average 99 stars, based on 572 article reviews
    Price from $9.99 to $1999.99
    2 mercaptoethanol - by Bioz Stars, 2020-02
    99/100 stars

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    Transduction:

    Article Title: A Myc-driven self-reinforcing regulatory network maintains mouse embryonic stem cell identity
    Article Snippet: Cell culture conditions R1 ESCs (obtained from American Type Culture Collection) were cultured without feeders on plastic coated with 0.1% gelatin in DMEM supplemented with 15% FCS (Hyclone Millipore, cat. ES-009-B), 100 mM 2-mercaptoethanol (Sigma, cat. M7522), 1 × MEM non-essential amino acids (Invitrogen, cat. 1140-036), 2 mM L -glutamine, 1 mM sodium pyruvate (Invitrogen), 100 μg ml−1 Vitamin C (L -ascorbic acid 2-phosphatase Sigma, A8960) and 100 U ml−1 LIF (Millipore, cat. ESG1107). .. The double-knock down for c- and N-Myc was obtained by lentivirus transduction followed by puromycin selection of MycER ESCs that were maintained in the presence of 50 nM OHT (Sigma, cat. H6278) and LIF.

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer. .. After the membranes were destained, they were blocked with 5% milk buffer, and washed and incubated overnight at 4 °C with 1:750 mouse anti-occludin monoclonal antibody (BD Transduction Laboratories, Franklin Lakes, NJ, USA) in 5% milk buffer with Tween 20.

    Clone Assay:

    Article Title: Control of directionality of chromatin folding for the inter- and intra-domain contacts at the Tfap2c–Bmp7 locus
    Article Snippet: The culture medium was DMEM (SIGMA-ALDRICH, Cat. D5796) containing 0.1 mM 2-mercaptoethanol (Sigma, Cat. M7522), leukemia inhibitory factor (Wako, Cat. 129-05601), penicillin–streptomycin–glutamine (Thermo Fisher Scientific, Cat. 10378-016), nonessential amino acids (Thermo Fisher Scientific, Cat. 11140-050) and 20% knockout serum replacement (Thermo Fisher Scientific, Cat. 10828-028). .. To perform the genome editing, we cloned the target sequences of CRISPR into the cloning site of pSpCas9(BB)-2A-Puro (PX459), which was gifted from Dr. Feng Zhang (Addgene plasmid # 48139), with Bbs I restriction enzyme [ ].

    Incubation:

    Article Title: Development of a novel in vivo corneal fibrosis model in the dog
    Article Snippet: Next, fixed tissues were rinsed with 100 mM sodium cacodylate buffer, pH 7.35 containing 10 mM 2-mercaptoethanol (Sigma Aldrich, St. Louis, MO, USA) and 130 mM sucrose (further referred to as 2-ME buffer). .. Specimens were next incubated at 4 °C for 1 hour and then rinsed with 2-ME buffer, which was followed by distilled water.

    Article Title: Caenorhabditis elegans Fibroblast Growth Factor Receptor Signaling Can Occur Independently of the Multi-Substrate Adaptor FRS2
    Article Snippet: .. An OD600 was taken, and the pellet of 1.0 ml of the overnight culture was resuspended in 1.0 ml of Z buffer (8.52 g anhydrous Na2 HPO4, 4.8 g anhydrous NaH2 PO4 , 0.12 g anhydrous MgSO4 , 0.74 g KCl dissolved in 1.0 liter water, and pH was adjusted to 7.0 with HCl), pelleted, and resuspended in 150 μl Z buffer supplemented with 2-mercaptoethanol (27 μl of 2-mercaptoethanol/10 ml Z buffer); 50 μl of chloroform and 20 μl of 0.1% SDS were added, and the sample was vortexed for 15 sec. A total of 700 μl of prewarmed (30°) ONPG (ortho-nitrophenyl-B(beta)-galactoside; 1 mg/ml in Z buffer with 2-mercaptoethanol; Sigma) was added, and the reactions were incubated at 30° until the solutions turned a yellow color. ..

    Article Title: Identification of new autoantibody specificities directed at proteins involved in the transforming growth factor ? pathway in patients with systemic sclerosis
    Article Snippet: Cells were then resuspended, incubated for 15 minutes on ice and regularly vortexed in a buffer containing 20 mM HEPES, pH 7.9, 0.4 M NaCl, 1 mM EDTA, 1 mM EGTA, 1 mM dithiothreitol and antiproteases. .. After ultracentrifugation, the supernatant was washed in a precooled (-20°C) solution of 10% trichloroacetic acid in acetone with 0.07% 2-mercaptoethanol (Sigma-Aldrich) to eliminate salts as described previously [ ].

    Article Title: Morphometric analysis of young petiole galls on the narrow-leaf cottonwood, Populus angustifolia, by the sugarbeet root aphid, Pemphigus betae
    Article Snippet: Tissues were fixed in 5 % glutaraldehyde in 50 mM sodium phosphate solution (pH 7), then rinsed with 100 mM sodium cacodylate buffer (pH 7.35) containing 10 mM 2-mercaptoethanol (Sigma Aldrich, St. Louis MO, USA) and 130 mM sucrose (further referred to as 2-ME buffer). .. Specimens were then incubated at 4 °C for 1 h, rinsed with 2-ME buffer followed by distilled water.

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer. .. After the membranes were destained, they were blocked with 5% milk buffer, and washed and incubated overnight at 4 °C with 1:750 mouse anti-occludin monoclonal antibody (BD Transduction Laboratories, Franklin Lakes, NJ, USA) in 5% milk buffer with Tween 20.

    Expressing:

    Article Title: Negative feedback via RSK modulates Erk‐dependent progression from naïve pluripotency
    Article Snippet: Zfp42 expression is associated with the naïve state and quickly downregulated as cells lose the capacity to self‐renew in 2iLIF conditions . .. Cells were grown in either GMEM medium (Sigma, cat. G5154) containing 15% FCS (Sigma, cat. F7524), 100 mM 2‐mercaptoethanol (Sigma, cat. M7522), 1× MEM non‐essential amino acids (Invitrogen, cat. 1140‐036), 2 mM l ‐glutamine, 1 mM sodium pyruvate (both from Invitrogen) and 100 units/ml LIF prepared in‐house, or in serum‐free N2B27 (prepared in‐house, or NDiff N2B27 base medium, Stem Cell Sciences Ltd, cat. SCS‐SF‐NB‐02, or NDiff 227 base medium, Takara Bio, cat. Y40002) supplemented with small molecule inhibitors PD (1 μM, PD0325901), CH (3 μM, CHIR99021) and LIF.

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: Paragraph title: 2.5. Evaluation of Occludin Protein Expression via Western Blot ... The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer.

    BIA-KA:

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer. .. The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer.

    Modification:

    Article Title: Fabrication of synthetic polymer coatings and their use in feeder-free culture of human embryonic stem cells
    Article Snippet: CHB-8 cells (Children's Hospital Corporation, cat. no. NIHhESC-09-0007) CHB-10 cells (Children's Hospital Corporation, cat. no. NIHhESC-09-0009) WA09 (H9) cells (WiCell Research Institute, cat. no.NIHhESC-10-0062) WA07 (H7) cells (WiCell Research Institute, cat. no. NIHhESC-10-0061) BG01 cells (BresaGen) Irradiated CF-1 mouse embryonic fibroblasts (MEF, passage 3; GlobalStem, cat. no. GSC-6001G) Gelatin type A, porcine (Sigma, cat. no. G1890; see REAGENT SETUP) Matrigel hESC-qualified Matrix (BD Biosciences, cat. no. 354277; see Box 1 ) Water (Sigma, cat. no. W3500) Dulbecco's phosphate buffered saline (D-PBS, without Ca2+ or Mg2+ ; GIBCO, cat. no. 14190) Dulbecco's modified Eagle medium (DMEM, high glucose; GIBCO, cat. no. 11965) DMEM/F12 (with l -glutamine and 15 mM HEPES; GIBCO, cat. no. 11330) KnockOut Serum Replacement (KOSR; GIBCO, cat. no. 10828) ▲ CRITICAL Store aliquots of KOSR at − 20 °C. .. Heat-inactivated FBS (HI-FBS; GIBCO, cat. no. 10082) l-glutamine (GIBCO, cat. no. 25030) Non-essential amino acids (GIBCO, cat. no. 11140) Penicillin/streptomycin (GIBCO, cat. no. 15140) BSA (GIBCO, cat. no. 15561) Trypan blue solution (Sigma, cat. no. T-8154) 2-Mercaptoethanol (Sigma, cat. no. M7522; see REAGENT SETUP) !

    Article Title: Ultrastructural Characteristics of Three Undifferentiated Mouse Embryonic Stem Cell Lines and Their differentiated Three-Dimensional Derivatives: A Comparative Study
    Article Snippet: The MEFs were grown in medium composed of Dulbecco's Modified Eagle Medium (DMEM; Gibco, cat. no. 41966052) supplemented with 10% fetal bovine serum (FBS; Gibco, cat. no. 26140087), 1% pen/strep (10,000 units of penicillin and 10,000 μg of streptomycin/mL; Gibco, cat. no. 15140122), and 1% nonessential amino acids (NEAA; X100; Gibco, cat. no. 11140035). .. The mESCs were cultured in medium composed of knockout D-MEM (Gibco, cat. no. 10829018) supplemented with 15% FBS-ES qualified (Gibco, cat. no. 16141079), 1% pen/strep (10,000 units of penicillin and 10,000 μg of streptomycin/mL; Gibco, cat. no. 15140122), 1% NEAA (X100; Gibco, cat. no. 11140035), 1% l -glutamine 200 mM (100×; Gibco, cat. no. 25030024), 1000 Units/mL of LIF (Millipore, cat. no. ESG1107), and 0.1 mM of 2-mercaptoethanol (Sigma, cat. no. M7522).

    Western Blot:

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: Paragraph title: 2.5. Evaluation of Occludin Protein Expression via Western Blot ... The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer.

    Derivative Assay:

    Article Title: Ultrastructural Characteristics of Three Undifferentiated Mouse Embryonic Stem Cell Lines and Their differentiated Three-Dimensional Derivatives: A Comparative Study
    Article Snippet: We used the mESCs derived from three different mouse strains, BALB/c, 129 W9.5, and C57BL/6, and propagated cells between passages 9 and 20. .. The mESCs were cultured in medium composed of knockout D-MEM (Gibco, cat. no. 10829018) supplemented with 15% FBS-ES qualified (Gibco, cat. no. 16141079), 1% pen/strep (10,000 units of penicillin and 10,000 μg of streptomycin/mL; Gibco, cat. no. 15140122), 1% NEAA (X100; Gibco, cat. no. 11140035), 1% l -glutamine 200 mM (100×; Gibco, cat. no. 25030024), 1000 Units/mL of LIF (Millipore, cat. no. ESG1107), and 0.1 mM of 2-mercaptoethanol (Sigma, cat. no. M7522).

    High Performance Liquid Chromatography:

    Article Title: Proteomic analysis of the medicinal plant Artemisia annua: Data from leaf and trichome extracts
    Article Snippet: 1.1 Solvents and solutions Solvents were of HPLC-grade and bought from Sigma-Aldrich, Poole, UK, except for water, which was purchased from Fisher Scientific, Loughborough, UK. .. The precipitation solution consisted of 10% (w/v) trichloroacetic acid (Fiedel-de Haen, Buchs, Switzerland) and 0.07% (w/v) 2-mercaptoethanol (Sigma-Aldrich) in cold acetone (Sigma-Aldrich) while the rinsing solution contained 0.07% (w/v) 2-mercaptoethanol in cold acetone.

    Electron Microscopy:

    Article Title: Development of a novel in vivo corneal fibrosis model in the dog
    Article Snippet: All reagents for this study’s TEM were purchased from Electron Microscopy Sciences and all specimen preparation was performed at the Electron Microscopy Core, University of Missouri, Columbia, MO, USA. .. Next, fixed tissues were rinsed with 100 mM sodium cacodylate buffer, pH 7.35 containing 10 mM 2-mercaptoethanol (Sigma Aldrich, St. Louis, MO, USA) and 130 mM sucrose (further referred to as 2-ME buffer).

    Article Title: Morphometric analysis of young petiole galls on the narrow-leaf cottonwood, Populus angustifolia, by the sugarbeet root aphid, Pemphigus betae
    Article Snippet: Paragraph title: Transmission electron microscopy ... Tissues were fixed in 5 % glutaraldehyde in 50 mM sodium phosphate solution (pH 7), then rinsed with 100 mM sodium cacodylate buffer (pH 7.35) containing 10 mM 2-mercaptoethanol (Sigma Aldrich, St. Louis MO, USA) and 130 mM sucrose (further referred to as 2-ME buffer).

    Transfection:

    Article Title: A Myc-driven self-reinforcing regulatory network maintains mouse embryonic stem cell identity
    Article Snippet: Cell culture conditions R1 ESCs (obtained from American Type Culture Collection) were cultured without feeders on plastic coated with 0.1% gelatin in DMEM supplemented with 15% FCS (Hyclone Millipore, cat. ES-009-B), 100 mM 2-mercaptoethanol (Sigma, cat. M7522), 1 × MEM non-essential amino acids (Invitrogen, cat. 1140-036), 2 mM L -glutamine, 1 mM sodium pyruvate (Invitrogen), 100 μg ml−1 Vitamin C (L -ascorbic acid 2-phosphatase Sigma, A8960) and 100 U ml−1 LIF (Millipore, cat. ESG1107). .. The R1 MycER ESCs clone A2 was obtained upon transfection with pBABE-MycER vector followed by puromycin selection.

    Protein Concentration:

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer. .. The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer.

    Protease Inhibitor:

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: Samples were mechanically homogenized in homogenization buffer (10 mM Tris base (Fisher-Scientific) with 37.22 mg Ethylenediaminetetraacetic acid (EDTA; Sigma–Aldrich, St. Louis, MO, USA), diluted in 100 mL distilled water, pH 7) containing protease inhibitor cocktail III, Mammalian (Research Products International, Mount Prospect, IL, USA) on ice. .. The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer.

    Cell Culture:

    Article Title: Esrrb Is a Pivotal Target of the Gsk3/Tcf3 Axis Regulating Embryonic Stem Cell Self-Renewal
    Article Snippet: .. Cells were cultured either in the GMEM (Sigma, cat. G5154) supplemented with 10% FCS (Sigma, cat. F7524), 100 μM 2-mercaptoethanol (Sigma, cat. M7522), 1× MEM nonessential amino acids (Invitrogen, cat. 1140-036), 2 mM L-glutamine, 1 mM sodium pyruvate (both from Invitrogen), and 100 units/ml LIF, or in the serum-free media N2B27 (NDiff N2B27 base medium, Stem Cell Sciences Ltd, cat. SCS-SF- NB-02) supplemented, as indicated, with small-molecule inhibitors PD (1 μM, PD0325901) and CH (3 μM, CHIR99021) and LIF prepared in-house. .. Colony forming assays were carried out by plating 600 ESCs per well on plates coated with laminin (Sigma, cat. L2020).

    Article Title: Rbm46 regulates mouse embryonic stem cell differentiation by targeting β-Catenin mRNA for degradation
    Article Snippet: .. Cell culture E14Tg2a mouse ESCs were cultured in the DMEM supplemented with 15% (v/v) fetal calf serum (FCS; Hyclone, Logan, UT, www.hyclone.com ), 100 mM 2- mercaptoethanol (Sigma; Cat. No. M7522), nonessential amino acids (Gibco), 2 mM l-glutamine (Chemicon), 1 mM sodium pyruvate (Sigma), and 100 U/mL leukemia inhibitory factor (LIF). .. Plates were fixed and stained for alkaline phosphatase (Sigma; Cat. No. 86R-1KT) according to the manufacturer’s protocol.

    Article Title: Stat3 promotes mitochondrial transcription and oxidative respiration during maintenance and induction of naive pluripotency
    Article Snippet: .. Cells were cultured either in serum‐free N2B27‐based medium (DMEM/F12 and Neurobasal [both Life Technologies] in 1:1 ratio, 0.1 mM 2‐mercaptoethanol, 2 mM l ‐glutamine, 1:200 N2 [Life Technologies], and 1:100 B27 [Life Technologies]) supplemented with small‐molecule inhibitors PD (1 μM, PD0325901), CH (3 mM, CHIR99021) from Axon (cat. 1386 and 1408) and LIF (100 units/ml produced in house), or in GMEM (Sigma, cat. G5154) supplemented with 10% FBS (Sigma, cat. F7524), 100 mM 2‐mercaptoethanol (Sigma, cat. M7522), 1× MEM non‐essential amino acids (Invitrogen, cat. 1140‐036), 2 mM l ‐glutamine, 1 mM sodium pyruvate (both from Invitrogen), and 100 units/ml LIF. .. EpiSCs were cultured without feeders on plastic coated with fibronectin (Millipore, cat. FC010) and replated every 2 days at a split ratio of 1:10 following dissociation with Dispase (Stem Cell Technologies, cat. 07923).

    Article Title: A Myc-driven self-reinforcing regulatory network maintains mouse embryonic stem cell identity
    Article Snippet: .. Cell culture conditions R1 ESCs (obtained from American Type Culture Collection) were cultured without feeders on plastic coated with 0.1% gelatin in DMEM supplemented with 15% FCS (Hyclone Millipore, cat. ES-009-B), 100 mM 2-mercaptoethanol (Sigma, cat. M7522), 1 × MEM non-essential amino acids (Invitrogen, cat. 1140-036), 2 mM L -glutamine, 1 mM sodium pyruvate (Invitrogen), 100 μg ml−1 Vitamin C (L -ascorbic acid 2-phosphatase Sigma, A8960) and 100 U ml−1 LIF (Millipore, cat. ESG1107). ..

    Article Title: Negative feedback via RSK modulates Erk‐dependent progression from naïve pluripotency
    Article Snippet: Paragraph title: ES cell culture ... Cells were grown in either GMEM medium (Sigma, cat. G5154) containing 15% FCS (Sigma, cat. F7524), 100 mM 2‐mercaptoethanol (Sigma, cat. M7522), 1× MEM non‐essential amino acids (Invitrogen, cat. 1140‐036), 2 mM l ‐glutamine, 1 mM sodium pyruvate (both from Invitrogen) and 100 units/ml LIF prepared in‐house, or in serum‐free N2B27 (prepared in‐house, or NDiff N2B27 base medium, Stem Cell Sciences Ltd, cat. SCS‐SF‐NB‐02, or NDiff 227 base medium, Takara Bio, cat. Y40002) supplemented with small molecule inhibitors PD (1 μM, PD0325901), CH (3 μM, CHIR99021) and LIF.

    Article Title: A Model-Based Analysis of Culture-Dependent Phenotypes of mESCs
    Article Snippet: .. Mouse embryonic stem cell culture Rex1GFPd2 embryonic stem cells (described in ) were cultured without feeders on plastic coated with 0.1% gelatine either in LIF/serum conditions (GMEM (Sigma, cat. G5154) supplemented with 10% FCS (Sigma, cat. F7524), 100 mM 2-mercaptoethanol (Sigma, cat. M7522), 13 MEM nonessential amino acids (Invitrogen, cat. 1140-036), 2 mM L-glutamine, 1 mM sodium pyruvate (both from Invitrogen), and 100 units/ml LIF), or in the serum-free media N2B27 (NDiff N2B27 base medium, Stem Cell Sciences Ltd, cat. SCS-SF-NB-02) supplemented with small-molecule inhibitors PD (1 mM, PD0325901) and CH (3 mM, CHIR99021) as described . .. Flow Cytometry After treatment with Accutase, live mESCs were resuspended in PBS with 1% FCS and ToPro-3 (Invitrogen) was added at a concentration of 0.05 nM to detect dead cells.

    Article Title: Control of directionality of chromatin folding for the inter- and intra-domain contacts at the Tfap2c–Bmp7 locus
    Article Snippet: Paragraph title: Cell culture and CRISPR genome editing ... The culture medium was DMEM (SIGMA-ALDRICH, Cat. D5796) containing 0.1 mM 2-mercaptoethanol (Sigma, Cat. M7522), leukemia inhibitory factor (Wako, Cat. 129-05601), penicillin–streptomycin–glutamine (Thermo Fisher Scientific, Cat. 10378-016), nonessential amino acids (Thermo Fisher Scientific, Cat. 11140-050) and 20% knockout serum replacement (Thermo Fisher Scientific, Cat. 10828-028).

    Article Title: Ultrastructural Characteristics of Three Undifferentiated Mouse Embryonic Stem Cell Lines and Their differentiated Three-Dimensional Derivatives: A Comparative Study
    Article Snippet: .. The mESCs were cultured in medium composed of knockout D-MEM (Gibco, cat. no. 10829018) supplemented with 15% FBS-ES qualified (Gibco, cat. no. 16141079), 1% pen/strep (10,000 units of penicillin and 10,000 μg of streptomycin/mL; Gibco, cat. no. 15140122), 1% NEAA (X100; Gibco, cat. no. 11140035), 1% l -glutamine 200 mM (100×; Gibco, cat. no. 25030024), 1000 Units/mL of LIF (Millipore, cat. no. ESG1107), and 0.1 mM of 2-mercaptoethanol (Sigma, cat. no. M7522). ..

    Transmission Assay:

    Article Title: Morphometric analysis of young petiole galls on the narrow-leaf cottonwood, Populus angustifolia, by the sugarbeet root aphid, Pemphigus betae
    Article Snippet: Paragraph title: Transmission electron microscopy ... Tissues were fixed in 5 % glutaraldehyde in 50 mM sodium phosphate solution (pH 7), then rinsed with 100 mM sodium cacodylate buffer (pH 7.35) containing 10 mM 2-mercaptoethanol (Sigma Aldrich, St. Louis MO, USA) and 130 mM sucrose (further referred to as 2-ME buffer).

    Sequencing:

    Article Title: Control of directionality of chromatin folding for the inter- and intra-domain contacts at the Tfap2c–Bmp7 locus
    Article Snippet: The culture medium was DMEM (SIGMA-ALDRICH, Cat. D5796) containing 0.1 mM 2-mercaptoethanol (Sigma, Cat. M7522), leukemia inhibitory factor (Wako, Cat. 129-05601), penicillin–streptomycin–glutamine (Thermo Fisher Scientific, Cat. 10378-016), nonessential amino acids (Thermo Fisher Scientific, Cat. 11140-050) and 20% knockout serum replacement (Thermo Fisher Scientific, Cat. 10828-028). .. The CRISPR target sequences and Oligo DNAs used to integrate the target sequence into the vector are listed in Additional file : Table S2.

    Transmission Electron Microscopy:

    Article Title: Development of a novel in vivo corneal fibrosis model in the dog
    Article Snippet: Paragraph title: 2.9. TEM Sample Preparation and Image Capture ... Next, fixed tissues were rinsed with 100 mM sodium cacodylate buffer, pH 7.35 containing 10 mM 2-mercaptoethanol (Sigma Aldrich, St. Louis, MO, USA) and 130 mM sucrose (further referred to as 2-ME buffer).

    Article Title: Morphometric analysis of young petiole galls on the narrow-leaf cottonwood, Populus angustifolia, by the sugarbeet root aphid, Pemphigus betae
    Article Snippet: Transmission electron microscopy Leaf samples (N = 5 for controls and N = 8 for galls) were processed for transmission electron microscopy (TEM) at the Electron Microscopy Core Facility (University of Missouri, Columbia MO, USA). .. Tissues were fixed in 5 % glutaraldehyde in 50 mM sodium phosphate solution (pH 7), then rinsed with 100 mM sodium cacodylate buffer (pH 7.35) containing 10 mM 2-mercaptoethanol (Sigma Aldrich, St. Louis MO, USA) and 130 mM sucrose (further referred to as 2-ME buffer).

    Isolation:

    Article Title: Proteomic analysis of the medicinal plant Artemisia annua: Data from leaf and trichome extracts
    Article Snippet: The isolation buffer contained 200 mM sorbitol (Fluka Biochemika, Buchs, Switzerland), 2 mM sucrose (Sigma-Aldrich), 5 mM succinic acid (Sigma-Aldrich), 5 mM dithiothreitol (Sigma-Aldrich), 1 mM ethylene glycol bis(2-aminoethyl ether)-N,N,N׳,N׳-tetraacetic acid (Sigma-Aldrich), 0.5 mM Na2 HPO4 (Sigma-Aldrich), 0.1 mM Na4 P2 O7 (Sigma-Aldrich), 25 mM HEPES (Sigma-Aldrich) and 5 mM MgCl2 (Sigma-Aldrich) in water. .. The precipitation solution consisted of 10% (w/v) trichloroacetic acid (Fiedel-de Haen, Buchs, Switzerland) and 0.07% (w/v) 2-mercaptoethanol (Sigma-Aldrich) in cold acetone (Sigma-Aldrich) while the rinsing solution contained 0.07% (w/v) 2-mercaptoethanol in cold acetone.

    Bicinchoninic Acid Protein Assay:

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer. .. The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer.

    Size-exclusion Chromatography:

    Article Title: Caenorhabditis elegans Fibroblast Growth Factor Receptor Signaling Can Occur Independently of the Multi-Substrate Adaptor FRS2
    Article Snippet: .. An OD600 was taken, and the pellet of 1.0 ml of the overnight culture was resuspended in 1.0 ml of Z buffer (8.52 g anhydrous Na2 HPO4, 4.8 g anhydrous NaH2 PO4 , 0.12 g anhydrous MgSO4 , 0.74 g KCl dissolved in 1.0 liter water, and pH was adjusted to 7.0 with HCl), pelleted, and resuspended in 150 μl Z buffer supplemented with 2-mercaptoethanol (27 μl of 2-mercaptoethanol/10 ml Z buffer); 50 μl of chloroform and 20 μl of 0.1% SDS were added, and the sample was vortexed for 15 sec. A total of 700 μl of prewarmed (30°) ONPG (ortho-nitrophenyl-B(beta)-galactoside; 1 mg/ml in Z buffer with 2-mercaptoethanol; Sigma) was added, and the reactions were incubated at 30° until the solutions turned a yellow color. ..

    Protein Extraction:

    Article Title: Identification of new autoantibody specificities directed at proteins involved in the transforming growth factor ? pathway in patients with systemic sclerosis
    Article Snippet: Paragraph title: Protein extraction ... After ultracentrifugation, the supernatant was washed in a precooled (-20°C) solution of 10% trichloroacetic acid in acetone with 0.07% 2-mercaptoethanol (Sigma-Aldrich) to eliminate salts as described previously [ ].

    Metabolic Labelling:

    Article Title: Negative feedback via RSK modulates Erk‐dependent progression from naïve pluripotency
    Article Snippet: Cells were grown in either GMEM medium (Sigma, cat. G5154) containing 15% FCS (Sigma, cat. F7524), 100 mM 2‐mercaptoethanol (Sigma, cat. M7522), 1× MEM non‐essential amino acids (Invitrogen, cat. 1140‐036), 2 mM l ‐glutamine, 1 mM sodium pyruvate (both from Invitrogen) and 100 units/ml LIF prepared in‐house, or in serum‐free N2B27 (prepared in‐house, or NDiff N2B27 base medium, Stem Cell Sciences Ltd, cat. SCS‐SF‐NB‐02, or NDiff 227 base medium, Takara Bio, cat. Y40002) supplemented with small molecule inhibitors PD (1 μM, PD0325901), CH (3 μM, CHIR99021) and LIF. .. For metabolic labelling, cells were cultured in arginine‐ and lysine‐free DMEM/F12 (Dundee Cell Products) supplemented with B27 (Gibco cat. 17504‐044), in‐house prepared N2 , 100 mM 2‐mercaptoethanol, 2 mM l ‐glutamine, 0.7 mM l ‐arginine (Sigma) or 13 C6 l ‐arginine (CK Gas Products), 0.5 mM l ‐lysine (Sigma) or 13 C6 l ‐lysine (CK Gas Products) and the two inhibitors PD and CH (2i) plus LIF.

    CRISPR:

    Article Title: Control of directionality of chromatin folding for the inter- and intra-domain contacts at the Tfap2c–Bmp7 locus
    Article Snippet: Paragraph title: Cell culture and CRISPR genome editing ... The culture medium was DMEM (SIGMA-ALDRICH, Cat. D5796) containing 0.1 mM 2-mercaptoethanol (Sigma, Cat. M7522), leukemia inhibitory factor (Wako, Cat. 129-05601), penicillin–streptomycin–glutamine (Thermo Fisher Scientific, Cat. 10378-016), nonessential amino acids (Thermo Fisher Scientific, Cat. 11140-050) and 20% knockout serum replacement (Thermo Fisher Scientific, Cat. 10828-028).

    Plasmid Preparation:

    Article Title: A Myc-driven self-reinforcing regulatory network maintains mouse embryonic stem cell identity
    Article Snippet: Cell culture conditions R1 ESCs (obtained from American Type Culture Collection) were cultured without feeders on plastic coated with 0.1% gelatin in DMEM supplemented with 15% FCS (Hyclone Millipore, cat. ES-009-B), 100 mM 2-mercaptoethanol (Sigma, cat. M7522), 1 × MEM non-essential amino acids (Invitrogen, cat. 1140-036), 2 mM L -glutamine, 1 mM sodium pyruvate (Invitrogen), 100 μg ml−1 Vitamin C (L -ascorbic acid 2-phosphatase Sigma, A8960) and 100 U ml−1 LIF (Millipore, cat. ESG1107). .. The R1 MycER ESCs clone A2 was obtained upon transfection with pBABE-MycER vector followed by puromycin selection.

    Article Title: Control of directionality of chromatin folding for the inter- and intra-domain contacts at the Tfap2c–Bmp7 locus
    Article Snippet: The culture medium was DMEM (SIGMA-ALDRICH, Cat. D5796) containing 0.1 mM 2-mercaptoethanol (Sigma, Cat. M7522), leukemia inhibitory factor (Wako, Cat. 129-05601), penicillin–streptomycin–glutamine (Thermo Fisher Scientific, Cat. 10378-016), nonessential amino acids (Thermo Fisher Scientific, Cat. 11140-050) and 20% knockout serum replacement (Thermo Fisher Scientific, Cat. 10828-028). .. To perform the genome editing, we cloned the target sequences of CRISPR into the cloning site of pSpCas9(BB)-2A-Puro (PX459), which was gifted from Dr. Feng Zhang (Addgene plasmid # 48139), with Bbs I restriction enzyme [ ].

    Irradiation:

    Article Title: Fabrication of synthetic polymer coatings and their use in feeder-free culture of human embryonic stem cells
    Article Snippet: CHB-8 cells (Children's Hospital Corporation, cat. no. NIHhESC-09-0007) CHB-10 cells (Children's Hospital Corporation, cat. no. NIHhESC-09-0009) WA09 (H9) cells (WiCell Research Institute, cat. no.NIHhESC-10-0062) WA07 (H7) cells (WiCell Research Institute, cat. no. NIHhESC-10-0061) BG01 cells (BresaGen) Irradiated CF-1 mouse embryonic fibroblasts (MEF, passage 3; GlobalStem, cat. no. GSC-6001G) Gelatin type A, porcine (Sigma, cat. no. G1890; see REAGENT SETUP) Matrigel hESC-qualified Matrix (BD Biosciences, cat. no. 354277; see Box 1 ) Water (Sigma, cat. no. W3500) Dulbecco's phosphate buffered saline (D-PBS, without Ca2+ or Mg2+ ; GIBCO, cat. no. 14190) Dulbecco's modified Eagle medium (DMEM, high glucose; GIBCO, cat. no. 11965) DMEM/F12 (with l -glutamine and 15 mM HEPES; GIBCO, cat. no. 11330) KnockOut Serum Replacement (KOSR; GIBCO, cat. no. 10828) ▲ CRITICAL Store aliquots of KOSR at − 20 °C. .. Heat-inactivated FBS (HI-FBS; GIBCO, cat. no. 10082) l-glutamine (GIBCO, cat. no. 25030) Non-essential amino acids (GIBCO, cat. no. 11140) Penicillin/streptomycin (GIBCO, cat. no. 15140) BSA (GIBCO, cat. no. 15561) Trypan blue solution (Sigma, cat. no. T-8154) 2-Mercaptoethanol (Sigma, cat. no. M7522; see REAGENT SETUP) !

    Stem Cell Culture:

    Article Title: Esrrb Is a Pivotal Target of the Gsk3/Tcf3 Axis Regulating Embryonic Stem Cell Self-Renewal
    Article Snippet: Paragraph title: Embryonic Stem Cell Culture ... Cells were cultured either in the GMEM (Sigma, cat. G5154) supplemented with 10% FCS (Sigma, cat. F7524), 100 μM 2-mercaptoethanol (Sigma, cat. M7522), 1× MEM nonessential amino acids (Invitrogen, cat. 1140-036), 2 mM L-glutamine, 1 mM sodium pyruvate (both from Invitrogen), and 100 units/ml LIF, or in the serum-free media N2B27 (NDiff N2B27 base medium, Stem Cell Sciences Ltd, cat. SCS-SF- NB-02) supplemented, as indicated, with small-molecule inhibitors PD (1 μM, PD0325901) and CH (3 μM, CHIR99021) and LIF prepared in-house.

    Article Title: Stat3 promotes mitochondrial transcription and oxidative respiration during maintenance and induction of naive pluripotency
    Article Snippet: Paragraph title: Embryonic stem cell culture ... Cells were cultured either in serum‐free N2B27‐based medium (DMEM/F12 and Neurobasal [both Life Technologies] in 1:1 ratio, 0.1 mM 2‐mercaptoethanol, 2 mM l ‐glutamine, 1:200 N2 [Life Technologies], and 1:100 B27 [Life Technologies]) supplemented with small‐molecule inhibitors PD (1 μM, PD0325901), CH (3 mM, CHIR99021) from Axon (cat. 1386 and 1408) and LIF (100 units/ml produced in house), or in GMEM (Sigma, cat. G5154) supplemented with 10% FBS (Sigma, cat. F7524), 100 mM 2‐mercaptoethanol (Sigma, cat. M7522), 1× MEM non‐essential amino acids (Invitrogen, cat. 1140‐036), 2 mM l ‐glutamine, 1 mM sodium pyruvate (both from Invitrogen), and 100 units/ml LIF.

    Article Title: A Model-Based Analysis of Culture-Dependent Phenotypes of mESCs
    Article Snippet: .. Mouse embryonic stem cell culture Rex1GFPd2 embryonic stem cells (described in ) were cultured without feeders on plastic coated with 0.1% gelatine either in LIF/serum conditions (GMEM (Sigma, cat. G5154) supplemented with 10% FCS (Sigma, cat. F7524), 100 mM 2-mercaptoethanol (Sigma, cat. M7522), 13 MEM nonessential amino acids (Invitrogen, cat. 1140-036), 2 mM L-glutamine, 1 mM sodium pyruvate (both from Invitrogen), and 100 units/ml LIF), or in the serum-free media N2B27 (NDiff N2B27 base medium, Stem Cell Sciences Ltd, cat. SCS-SF-NB-02) supplemented with small-molecule inhibitors PD (1 mM, PD0325901) and CH (3 mM, CHIR99021) as described . .. Flow Cytometry After treatment with Accutase, live mESCs were resuspended in PBS with 1% FCS and ToPro-3 (Invitrogen) was added at a concentration of 0.05 nM to detect dead cells.

    Selection:

    Article Title: A Myc-driven self-reinforcing regulatory network maintains mouse embryonic stem cell identity
    Article Snippet: Cell culture conditions R1 ESCs (obtained from American Type Culture Collection) were cultured without feeders on plastic coated with 0.1% gelatin in DMEM supplemented with 15% FCS (Hyclone Millipore, cat. ES-009-B), 100 mM 2-mercaptoethanol (Sigma, cat. M7522), 1 × MEM non-essential amino acids (Invitrogen, cat. 1140-036), 2 mM L -glutamine, 1 mM sodium pyruvate (Invitrogen), 100 μg ml−1 Vitamin C (L -ascorbic acid 2-phosphatase Sigma, A8960) and 100 U ml−1 LIF (Millipore, cat. ESG1107). .. The R1 MycER ESCs clone A2 was obtained upon transfection with pBABE-MycER vector followed by puromycin selection.

    Sample Prep:

    Article Title: Development of a novel in vivo corneal fibrosis model in the dog
    Article Snippet: Paragraph title: 2.9. TEM Sample Preparation and Image Capture ... Next, fixed tissues were rinsed with 100 mM sodium cacodylate buffer, pH 7.35 containing 10 mM 2-mercaptoethanol (Sigma Aldrich, St. Louis, MO, USA) and 130 mM sucrose (further referred to as 2-ME buffer).

    Homogenization:

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: Samples were mechanically homogenized in homogenization buffer (10 mM Tris base (Fisher-Scientific) with 37.22 mg Ethylenediaminetetraacetic acid (EDTA; Sigma–Aldrich, St. Louis, MO, USA), diluted in 100 mL distilled water, pH 7) containing protease inhibitor cocktail III, Mammalian (Research Products International, Mount Prospect, IL, USA) on ice. .. The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer.

    Knock-Out:

    Article Title: Fabrication of synthetic polymer coatings and their use in feeder-free culture of human embryonic stem cells
    Article Snippet: CHB-8 cells (Children's Hospital Corporation, cat. no. NIHhESC-09-0007) CHB-10 cells (Children's Hospital Corporation, cat. no. NIHhESC-09-0009) WA09 (H9) cells (WiCell Research Institute, cat. no.NIHhESC-10-0062) WA07 (H7) cells (WiCell Research Institute, cat. no. NIHhESC-10-0061) BG01 cells (BresaGen) Irradiated CF-1 mouse embryonic fibroblasts (MEF, passage 3; GlobalStem, cat. no. GSC-6001G) Gelatin type A, porcine (Sigma, cat. no. G1890; see REAGENT SETUP) Matrigel hESC-qualified Matrix (BD Biosciences, cat. no. 354277; see Box 1 ) Water (Sigma, cat. no. W3500) Dulbecco's phosphate buffered saline (D-PBS, without Ca2+ or Mg2+ ; GIBCO, cat. no. 14190) Dulbecco's modified Eagle medium (DMEM, high glucose; GIBCO, cat. no. 11965) DMEM/F12 (with l -glutamine and 15 mM HEPES; GIBCO, cat. no. 11330) KnockOut Serum Replacement (KOSR; GIBCO, cat. no. 10828) ▲ CRITICAL Store aliquots of KOSR at − 20 °C. .. Heat-inactivated FBS (HI-FBS; GIBCO, cat. no. 10082) l-glutamine (GIBCO, cat. no. 25030) Non-essential amino acids (GIBCO, cat. no. 11140) Penicillin/streptomycin (GIBCO, cat. no. 15140) BSA (GIBCO, cat. no. 15561) Trypan blue solution (Sigma, cat. no. T-8154) 2-Mercaptoethanol (Sigma, cat. no. M7522; see REAGENT SETUP) !

    Article Title: Control of directionality of chromatin folding for the inter- and intra-domain contacts at the Tfap2c–Bmp7 locus
    Article Snippet: .. The culture medium was DMEM (SIGMA-ALDRICH, Cat. D5796) containing 0.1 mM 2-mercaptoethanol (Sigma, Cat. M7522), leukemia inhibitory factor (Wako, Cat. 129-05601), penicillin–streptomycin–glutamine (Thermo Fisher Scientific, Cat. 10378-016), nonessential amino acids (Thermo Fisher Scientific, Cat. 11140-050) and 20% knockout serum replacement (Thermo Fisher Scientific, Cat. 10828-028). .. We cultured the cells on the SNL feeder cells to maintain and on dish coated with thin layer of Matrigel (Corning, Cat. 354277) without feeder cells to expand for use for the 4C-seq and N-ChIP assays.

    Article Title: Ultrastructural Characteristics of Three Undifferentiated Mouse Embryonic Stem Cell Lines and Their differentiated Three-Dimensional Derivatives: A Comparative Study
    Article Snippet: .. The mESCs were cultured in medium composed of knockout D-MEM (Gibco, cat. no. 10829018) supplemented with 15% FBS-ES qualified (Gibco, cat. no. 16141079), 1% pen/strep (10,000 units of penicillin and 10,000 μg of streptomycin/mL; Gibco, cat. no. 15140122), 1% NEAA (X100; Gibco, cat. no. 11140035), 1% l -glutamine 200 mM (100×; Gibco, cat. no. 25030024), 1000 Units/mL of LIF (Millipore, cat. no. ESG1107), and 0.1 mM of 2-mercaptoethanol (Sigma, cat. no. M7522). ..

    Produced:

    Article Title: Stat3 promotes mitochondrial transcription and oxidative respiration during maintenance and induction of naive pluripotency
    Article Snippet: .. Cells were cultured either in serum‐free N2B27‐based medium (DMEM/F12 and Neurobasal [both Life Technologies] in 1:1 ratio, 0.1 mM 2‐mercaptoethanol, 2 mM l ‐glutamine, 1:200 N2 [Life Technologies], and 1:100 B27 [Life Technologies]) supplemented with small‐molecule inhibitors PD (1 μM, PD0325901), CH (3 mM, CHIR99021) from Axon (cat. 1386 and 1408) and LIF (100 units/ml produced in house), or in GMEM (Sigma, cat. G5154) supplemented with 10% FBS (Sigma, cat. F7524), 100 mM 2‐mercaptoethanol (Sigma, cat. M7522), 1× MEM non‐essential amino acids (Invitrogen, cat. 1140‐036), 2 mM l ‐glutamine, 1 mM sodium pyruvate (both from Invitrogen), and 100 units/ml LIF. .. EpiSCs were cultured without feeders on plastic coated with fibronectin (Millipore, cat. FC010) and replated every 2 days at a split ratio of 1:10 following dissociation with Dispase (Stem Cell Technologies, cat. 07923).

    N-ChIP:

    Article Title: Control of directionality of chromatin folding for the inter- and intra-domain contacts at the Tfap2c–Bmp7 locus
    Article Snippet: The culture medium was DMEM (SIGMA-ALDRICH, Cat. D5796) containing 0.1 mM 2-mercaptoethanol (Sigma, Cat. M7522), leukemia inhibitory factor (Wako, Cat. 129-05601), penicillin–streptomycin–glutamine (Thermo Fisher Scientific, Cat. 10378-016), nonessential amino acids (Thermo Fisher Scientific, Cat. 11140-050) and 20% knockout serum replacement (Thermo Fisher Scientific, Cat. 10828-028). .. We cultured the cells on the SNL feeder cells to maintain and on dish coated with thin layer of Matrigel (Corning, Cat. 354277) without feeder cells to expand for use for the 4C-seq and N-ChIP assays.

    Staining:

    Article Title: Rbm46 regulates mouse embryonic stem cell differentiation by targeting β-Catenin mRNA for degradation
    Article Snippet: Cell culture E14Tg2a mouse ESCs were cultured in the DMEM supplemented with 15% (v/v) fetal calf serum (FCS; Hyclone, Logan, UT, www.hyclone.com ), 100 mM 2- mercaptoethanol (Sigma; Cat. No. M7522), nonessential amino acids (Gibco), 2 mM l-glutamine (Chemicon), 1 mM sodium pyruvate (Sigma), and 100 U/mL leukemia inhibitory factor (LIF). .. Plates were fixed and stained for alkaline phosphatase (Sigma; Cat. No. 86R-1KT) according to the manufacturer’s protocol.

    Article Title: Negative feedback via RSK modulates Erk‐dependent progression from naïve pluripotency
    Article Snippet: Cells were grown in either GMEM medium (Sigma, cat. G5154) containing 15% FCS (Sigma, cat. F7524), 100 mM 2‐mercaptoethanol (Sigma, cat. M7522), 1× MEM non‐essential amino acids (Invitrogen, cat. 1140‐036), 2 mM l ‐glutamine, 1 mM sodium pyruvate (both from Invitrogen) and 100 units/ml LIF prepared in‐house, or in serum‐free N2B27 (prepared in‐house, or NDiff N2B27 base medium, Stem Cell Sciences Ltd, cat. SCS‐SF‐NB‐02, or NDiff 227 base medium, Takara Bio, cat. Y40002) supplemented with small molecule inhibitors PD (1 μM, PD0325901), CH (3 μM, CHIR99021) and LIF. .. For colony formation assays, cells were plated on laminin‐coated plates in 2iLIF for 5 days and fixed and stained for alkaline phosphatase (Sigma, cat. 86 R‐1KT) according to the manufacturer's instructions.

    Article Title: Peripheral Anti-Angiogenic Imbalance during Pregnancy Impairs Myogenic Tone and Increases Cerebral Edema in a Rodent Model of HELLP Syndrome
    Article Snippet: .. The samples were mixed with sample buffer (125 mM Tris base, 20% glycerol (Sigma–Aldrich,), 4% sodium dodecyl sulfate solution (SDS, Bio-Rad, Hercules, CA, USA), 10% mercaptoethanol (Sigma–Aldrich, St. Louis, MO, USA), 0.05% bromophenol blue (Sigma–Aldrich), pH 6.8) and heated at 95 °C for 10 min. Next, 50 µg of the proteins was electrophoresed (BioRad, Hercules, CA, USA) and transferred onto nitrocellulose membranes followed by immersion in Ponceau-S stain (Sigma–Aldrich, St. Louis, MO, USA)) to ensure protein transfer. .. After the membranes were destained, they were blocked with 5% milk buffer, and washed and incubated overnight at 4 °C with 1:750 mouse anti-occludin monoclonal antibody (BD Transduction Laboratories, Franklin Lakes, NJ, USA) in 5% milk buffer with Tween 20.

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  • 95
    Millipore cathepsin b inhibitor ca 074 me
    Pan-caspase inhibitor Z-VAD-FMK and the <t>cathepsin</t> B inhibitor <t>CA-074-Me</t> inhibit Methylbenzoprim-induced apoptosis in melanoma cells. The percentage of apoptotic cells was evaluated after 24, 48 and 72 h of MBP (0.8, 8 μg/ml), the positive control Pyr (8 μg/ml) and untreated cells in Mel501 (panel a ) and MeWo (panel b ) cell lines pretreated for 2 h with the pan-caspase inhibitor Z-VAD-FMK (50 μmol/L) or the cathepsin B inhibitor CA-074-Me (10 μmol/L) or both. Columns , mean values of three independent experiments; bars, SD. *, P
    Cathepsin B Inhibitor Ca 074 Me, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 3 article reviews
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    cathepsin b inhibitor ca 074 me - by Bioz Stars, 2020-02
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    79
    Millipore noc 7
    Depletion of kinesin-5 reduces the distance of axonal retraction. (A and B) DIC images of control axons (A) and kinesin-5–depleted axons (B) before and 30 min after treatment with 0.3 mM <t>noc-7.</t> (B) Kinesin-5–depleted axons continue to elongate even in the presence of noc-7. Arrows demarcate the distal tips of growth cones before and after noc-7 treatment. (C) Quantitative analysis of noc-7–induced axonal retraction revealed a statistically significant reduction in mean distance retracted in neurons depleted of kinesin-5 (mean ± SEM; control, n = 54; kinesin-5 siRNA, n = 55; *, P = 0.011). Bar, 20 μm.
    Noc 7, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore ksr medium
    LCD shows direct differentiation outcomes of <t>H9</t> cells seeded at different densities A-D , Undifferentiated H9 cells with localized high cell density were subjected to IF using anti-PAX6 (A and D) and anti-OCT4 (B and D) antibodies. E-L , H9 cells seeded at low density (8×10 3 cells/ cm 2 ) (E-H) and high density (1×10 4 cells/ cm 2 ) (I-L) were treated with <t>KSR</t> and N2 medium supplemented with noggin and SB431542 for 5 days. The cells were then subjected to the IF assay using anti-PAX6 antibody (green, E, H, I and L) and anti-OCT4 antibody (red, F, H, J and L) in the H9-derived cells. M, The areas of the signals in the OCT4, PAX6 and DAPI channels were calculated using Image J software. The ratios of the OCT4 and PAX6 area to DAPI area are shown (X±SD, n=8; *, P
    Ksr Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore laemmli buffer
    Immunoreactivity of antibody against rat SGLT2 (rSGLT2-Ab) in kidneys and small intestine of female rats. A : optimal conditions for Western blots with total cell membranes (TCM) from rat kidney cortex. For SDS-PAGE, isolated TCM were prepared in <t>Laemmli</t>
    Laemmli Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 92 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Pan-caspase inhibitor Z-VAD-FMK and the cathepsin B inhibitor CA-074-Me inhibit Methylbenzoprim-induced apoptosis in melanoma cells. The percentage of apoptotic cells was evaluated after 24, 48 and 72 h of MBP (0.8, 8 μg/ml), the positive control Pyr (8 μg/ml) and untreated cells in Mel501 (panel a ) and MeWo (panel b ) cell lines pretreated for 2 h with the pan-caspase inhibitor Z-VAD-FMK (50 μmol/L) or the cathepsin B inhibitor CA-074-Me (10 μmol/L) or both. Columns , mean values of three independent experiments; bars, SD. *, P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: New derivatives of the antimalarial drug Pyrimethamine in the control of melanoma tumor growth: an in vitro and in vivo study

    doi: 10.1186/s13046-016-0409-9

    Figure Lengend Snippet: Pan-caspase inhibitor Z-VAD-FMK and the cathepsin B inhibitor CA-074-Me inhibit Methylbenzoprim-induced apoptosis in melanoma cells. The percentage of apoptotic cells was evaluated after 24, 48 and 72 h of MBP (0.8, 8 μg/ml), the positive control Pyr (8 μg/ml) and untreated cells in Mel501 (panel a ) and MeWo (panel b ) cell lines pretreated for 2 h with the pan-caspase inhibitor Z-VAD-FMK (50 μmol/L) or the cathepsin B inhibitor CA-074-Me (10 μmol/L) or both. Columns , mean values of three independent experiments; bars, SD. *, P

    Article Snippet: The cathepsin B inhibitor CA-074-Me (Calbiochem, Millipore, Germany) and pan-caspase inhibitor z-VAD-fmk (R & D System, USA) were diluted in RPMI 1640 immediately before experiments.

    Techniques: Positive Control

    Depletion of kinesin-5 reduces the distance of axonal retraction. (A and B) DIC images of control axons (A) and kinesin-5–depleted axons (B) before and 30 min after treatment with 0.3 mM noc-7. (B) Kinesin-5–depleted axons continue to elongate even in the presence of noc-7. Arrows demarcate the distal tips of growth cones before and after noc-7 treatment. (C) Quantitative analysis of noc-7–induced axonal retraction revealed a statistically significant reduction in mean distance retracted in neurons depleted of kinesin-5 (mean ± SEM; control, n = 54; kinesin-5 siRNA, n = 55; *, P = 0.011). Bar, 20 μm.

    Journal: The Journal of Cell Biology

    Article Title: Kinesin-5 regulates the growth of the axon by acting as a brake on its microtubule array

    doi: 10.1083/jcb.200702074

    Figure Lengend Snippet: Depletion of kinesin-5 reduces the distance of axonal retraction. (A and B) DIC images of control axons (A) and kinesin-5–depleted axons (B) before and 30 min after treatment with 0.3 mM noc-7. (B) Kinesin-5–depleted axons continue to elongate even in the presence of noc-7. Arrows demarcate the distal tips of growth cones before and after noc-7 treatment. (C) Quantitative analysis of noc-7–induced axonal retraction revealed a statistically significant reduction in mean distance retracted in neurons depleted of kinesin-5 (mean ± SEM; control, n = 54; kinesin-5 siRNA, n = 55; *, P = 0.011). Bar, 20 μm.

    Article Snippet: DIC images of axons were recorded before and 30 min after addition of noc-7.

    Techniques:

    LCD shows direct differentiation outcomes of H9 cells seeded at different densities A-D , Undifferentiated H9 cells with localized high cell density were subjected to IF using anti-PAX6 (A and D) and anti-OCT4 (B and D) antibodies. E-L , H9 cells seeded at low density (8×10 3 cells/ cm 2 ) (E-H) and high density (1×10 4 cells/ cm 2 ) (I-L) were treated with KSR and N2 medium supplemented with noggin and SB431542 for 5 days. The cells were then subjected to the IF assay using anti-PAX6 antibody (green, E, H, I and L) and anti-OCT4 antibody (red, F, H, J and L) in the H9-derived cells. M, The areas of the signals in the OCT4, PAX6 and DAPI channels were calculated using Image J software. The ratios of the OCT4 and PAX6 area to DAPI area are shown (X±SD, n=8; *, P

    Journal: Biochemical and biophysical research communications

    Article Title: Synergistic contribution of SMAD signaling blockade and high localized cell density in the differentiation of neuroectoderm from H9 cells

    doi: 10.1016/j.bbrc.2014.08.137

    Figure Lengend Snippet: LCD shows direct differentiation outcomes of H9 cells seeded at different densities A-D , Undifferentiated H9 cells with localized high cell density were subjected to IF using anti-PAX6 (A and D) and anti-OCT4 (B and D) antibodies. E-L , H9 cells seeded at low density (8×10 3 cells/ cm 2 ) (E-H) and high density (1×10 4 cells/ cm 2 ) (I-L) were treated with KSR and N2 medium supplemented with noggin and SB431542 for 5 days. The cells were then subjected to the IF assay using anti-PAX6 antibody (green, E, H, I and L) and anti-OCT4 antibody (red, F, H, J and L) in the H9-derived cells. M, The areas of the signals in the OCT4, PAX6 and DAPI channels were calculated using Image J software. The ratios of the OCT4 and PAX6 area to DAPI area are shown (X±SD, n=8; *, P

    Article Snippet: Briefly, H9 cells were cultured on MEFs in KSR medium (DMEM/F12, 20 % KSR, 0.1 mM β-mercaptoethanol, 10 ng/ml of FGF-2) and disaggregated using accutase (Millipore, Billerica, MA, USA) for 20 min, washed with KSR medium and pre-plated on gelatin-coated 6-well plates for 1 h at 37 °C in the presence of the ROCK inhibitor (Y-27632) to remove MEFs.

    Techniques: Derivative Assay, Software

    Synergistic contribution of SMAD signaling blockers and localized high cell density in NE differentiation A-F, Five days after the cell-clump-based differentiation of NE in KSR and N2 medium with (D-F) or without (A-C) SMAD signaling blockers, H9-derived cells were subjected to the IF assay using anti-PAX6 antibody (green, A-D). The nuclei were stained using DAPI (blue, C and D). The micrographs were divided into 20 (5×4) squares as indicated (C and D). E-F , The number of total cells and PAX6-positive cells in each square was quantified using Image J software. The ratio of PAX6-positive cells to total cells in each square was determined. The squares with equivalent ratios were binned together. The ratios of PAX6-positive cells to total cells in each bin are shown (F, X±SD, **P

    Journal: Biochemical and biophysical research communications

    Article Title: Synergistic contribution of SMAD signaling blockade and high localized cell density in the differentiation of neuroectoderm from H9 cells

    doi: 10.1016/j.bbrc.2014.08.137

    Figure Lengend Snippet: Synergistic contribution of SMAD signaling blockers and localized high cell density in NE differentiation A-F, Five days after the cell-clump-based differentiation of NE in KSR and N2 medium with (D-F) or without (A-C) SMAD signaling blockers, H9-derived cells were subjected to the IF assay using anti-PAX6 antibody (green, A-D). The nuclei were stained using DAPI (blue, C and D). The micrographs were divided into 20 (5×4) squares as indicated (C and D). E-F , The number of total cells and PAX6-positive cells in each square was quantified using Image J software. The ratio of PAX6-positive cells to total cells in each square was determined. The squares with equivalent ratios were binned together. The ratios of PAX6-positive cells to total cells in each bin are shown (F, X±SD, **P

    Article Snippet: Briefly, H9 cells were cultured on MEFs in KSR medium (DMEM/F12, 20 % KSR, 0.1 mM β-mercaptoethanol, 10 ng/ml of FGF-2) and disaggregated using accutase (Millipore, Billerica, MA, USA) for 20 min, washed with KSR medium and pre-plated on gelatin-coated 6-well plates for 1 h at 37 °C in the presence of the ROCK inhibitor (Y-27632) to remove MEFs.

    Techniques: Derivative Assay, Staining, Software

    Immunoreactivity of antibody against rat SGLT2 (rSGLT2-Ab) in kidneys and small intestine of female rats. A : optimal conditions for Western blots with total cell membranes (TCM) from rat kidney cortex. For SDS-PAGE, isolated TCM were prepared in Laemmli

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Expression of Na+-d-glucose cotransporter SGLT2 in rodents is kidney-specific and exhibits sex and species differences

    doi: 10.1152/ajpcell.00450.2011

    Figure Lengend Snippet: Immunoreactivity of antibody against rat SGLT2 (rSGLT2-Ab) in kidneys and small intestine of female rats. A : optimal conditions for Western blots with total cell membranes (TCM) from rat kidney cortex. For SDS-PAGE, isolated TCM were prepared in Laemmli

    Article Snippet: Denaturation of membrane samples in Laemmli buffer [±β-mercaptoethanol (β-ME)] and heating at different temperatures (37°C for 30 min, 65°C for 15 min, 95°C for 5 min), separation of the proteins through 10% SDS-PAGE minigels, as well as electrophoretic wet transfer to an Immobilon membrane (Millipore, Bedford, MA) were performed as described previously ( , ).

    Techniques: Western Blot, SDS Page, Isolation