JIMT-1 (1.5 × 104/mL) and BT474 (5 × 104/mL) cells were plated using ultralow attachment dishes and grown in HuMEC basal serum free medium (Gibco) with B27 (1:50, Invitrogen), 20 ng/mL human epidermal growth factor (EGF, Sigma-Aldrich), 20 ng/mL basic fibroblast growth factor (bFGF, Sigma-Aldrich), 1% antibiotic–antimycotic, 4 μg/mL heparin, and 15 μg/mL gentamycin at 37 °C (5% CO2). The number and volumes of the mammospheres was assessed with a CKX53 inverted microscope (Olympus Life Science). Mammosphere volumes were calculated by the formula volume = 4/3*3.14(π)*r3 (r: radius).
Human epidermal growth factor egf
Manufactured by Merck Group
Sourced in United States
Human epidermal growth factor (EGF) is a protein that stimulates cell growth and differentiation. It plays a crucial role in the regulation of cell proliferation, survival, and migration. EGF binds to its receptor (EGFR) on the cell surface, activating intracellular signaling pathways that promote various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Humec basal serum free medium, by Thermo Fisher Scientific (4 mentions)
Basic fibroblast growth factor bfgf, by Merck Group (3 mentions)
Ckx53 inverted microscope, by Olympus (3 mentions)
Penicillin streptomycin, by Lonza (3 mentions)
Fetal bovine serum (fbs), by Lonza (3 mentions)
B27 supplement, by Thermo Fisher Scientific (3 mentions)
Hydrocortisone, by Merck Group (2 mentions)
Trypsin edta, by Lonza (2 mentions)
12 well ultra low adherent cell culture plate, by Corning (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Skim milk, by BD (1 mentions)
Chicken egg white albumin, by Merck Group (1 mentions)
Human insulin, by Merck Group (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Donkey anti goat igg, by Jackson ImmunoResearch (1 mentions)
Essential fatty acid free bovine serum albumin, by BD (1 mentions)
Muc5ac antibody, by Abcam (1 mentions)
Apo transferrin, by Merck Group (1 mentions)
β actin antibody, by Merck Group (1 mentions)
18 protocols using human epidermal growth factor egf
1
Mammosphere Formation Assay for JIMT-1 and BT474 Cells
2
Optimized Cell Culture Conditions
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M199, human epidermal growth factor (EGF), hydrocortisone, gelatin, human insulin, apotransferrin, chicken egg white albumin and LPS were obtained from Sigma-Aldrich Chemical (St. Louis, MO, USA), as were all other reagents, unless specifically stated otherwise. Fetal bovine serum (FBS), penicillin-streptomycin and trypsin-EDTA were purchased from Lonza (Walkersville, MD, USA). Eotaxin-1 (CCL11) antibody was purchased from R&D systems (Minneapolis, MN, USA) and β-actin antibody was obtained from Sigma-Aldrich Chemicals. Additionally, MUC5AC antibody was provided by Abcam (Cambridge, UK). Antibodies of toll-like receptor 4 (TLR4), intracellular adhesion molecule (ICAM)-1 and eosinophil major basic protein (EMBP) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG, donkey anti-goat IgG and goat anti-mouse IgG were purchased from Jackson Immuno-Research Laboratories (West Grove, PA, USA). Essential fatty acid free bovine serum albumin (BSA) and skim milk were supplied by Becton Dickinson Company (Sparks, MD, USA).
3
Mammosphere Formation Assay Protocol
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Cells were plated at a density of 2,000 cells/well in 12-well ultra-low adherent cell culture plate (Corning, USA) and grown in DMEM/F12 containing 1x B27 supplement (Invitrogen), 20 ng/mL human epidermal growth factor (EGF; Sigma) and 20 ng/ml of human basic fibroblast growth factor (bFGF; R&D Systems) for 7-14 days without disturbing the plate. After culturing, the mammospheres with diameter greater than 40 µm were counted under a microscope for quantitative analysis.
4
Enrichment and Quantification of Mammospheres
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BT474 (3.0×105) or JIMT-1 (1.5×105) cells were plated in ultralow attachment dishes and cultured in HuMEC basal serum-free medium (Gibco), supplemented with B27 (1:50, Invitrogen), 20 ng/mL basic fibroblast growth factor (bFGF, Sigma-Aldrich), 20 ng/mL human epidermal growth factor (EGF, Sigma-Aldrich), 4 μg/mL heparin, 1% antibiotic-antimycotic agent, and 15 μg/mL gentamycin. The numbers and volumes of mammospheres were determined under an Olympus CKX53 inverted microscope (Olympus Life Science). Mammosphere volumes were calculated by the formula: volume = 4/3*3.14(π)*r3 (r: radius).
5
Differentiation of ReNcell VM Neural Progenitors
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The immortalized human ReNcell VM (Ventral Mesencephalon) neural progenitor cells (EMD Millipore) were cultured on 20 μg/ml laminin-coated plates (Corning), in Dulbecco’s Modified Eagle Medium (DMEM): Nutrient Mixture F12 (DMEM/F12) media (Life Technologies) supplemented with 2% (v/v) B27 neural supplement (Life Technologies), 2 μg/ml heparin (StemCell Technologies), 20 μg/ml human Epidermal Growth Factor (EGF; Sigma), 20 μg/ml human Fibroblast Growth Factor-basic (bFGF; Life Technologies), and 100 units/ml penicillin-streptomycin solution (GE Hyclone). The culture media was changed every 3 days until the cells were confluent and ready for subculturing. Differentiation of the neural progenitor cells was initiated by replacing the culture media with DMEM/F12 supplemented with 2% (v/v) B27 neural supplement, 2 μg/ml heparin, and 100 units/ml penicillin-streptomycin solution without the growth factors EGF and bFGF. The differentiation media was changed every 3 days for 3–4 weeks. All cells were grown in 5% CO2 at 37 °C.
6
Mammosphere Formation Assay Protocol
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SUM-149 cells for mammosphere formation were cultured in Ham’s F-12 medium containing 20 ng/mL EGF (Sigma), 20 ng/mL basic FGF (Fisher Scientific), 1X B27 (Invitrogen), 4 ng/mL Heparin (Sigma), 5 µg/mL insulin, 1 µg/mL hydrocortisone, 100 units/mL penicillin and 100 units/mL streptomycin. The MDA-MB-231 were cultured in HuMEC basal serum-free medium (Gibco), supplemented with B27 (1:50, Invitrogen, Carlsbad, CA, USA), 20 ng/mL basic fibroblast growth factor (bFGF, Sigma-Aldrich), 20 ng/mL human epidermal growth factor (EGF, Sigma-Aldrich), 4 μg/mL Heparin, 1% antibiotic-antimycotic agent, and 15 μg/mL gentamycin. Cells were trypsinized, passed through a 40 µm cell strainer (BD Falcon), seeded in ultra-low attachment plates (Corning), cultured for 7–10 days at a density of 10,000 cells/mL for SUM-149 and 5,000 cells/mL for the MDA-MB-231 cells and the experiments were performed after the 3rd passage. The mammospheres were treated with vehicle or GLE for 24 h and analyzed using an inverted microscope. Photos were acquired with ZEN software from ZEISS Microscopy (Oberkochen, Germany). Circularity was calculated using the formula 4π (Area/Perimeter2) using Image J [59 (link)–61 (link), 67 (link)].
7
Bronchial Epithelial Cell Responses to Environmental Particulates
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M199 culture media and human epidermal growth factor (EGF) were obtained from the Sigma-Aldrich Chemical (St. Louis, MO, USA), as were all other reagents, unless specifically stated elsewhere. Fetal bovine serum (FBS), penicillin-streptomycin, and trypsin-EDTA were offered by Lonza (Walkersville, MD, USA). Human bronchial airway epithelial cell line, BEAS-2B, was provided from the American Type Culture Collection (ATCC, Manassas, VA, USA). Sigma-Aldrich Chemical provided uPM10 sample (product number: NIST SRM 1648A, particles less than 10 μm). For the Western blot analysis, antibodies against toll-like receptor 4 (TLR4), cyclooxygenase-2 (COX-2), nitric oxide synthase 2 (NOS2), eotaxin-1, α-smooth muscle actin (α-SMA), and collagen I were obtained from the Santa Cruz Biotechnology (Santa Cruz, CA, USA). Human collagen IV antibody was purchased from Bioss Antibodies (Woburn, MA, USA). Antibodies of human EGF receptor (EGFR) and MUC5AC were supplied by Abcam (Cambridge, UK).
Aesculetin (Sigma-Aldrich Chemical) was prepared in dimethyl sulfoxide (DMSO, <0.5% in a final culture concentration) for cell culture.
Aesculetin (Sigma-Aldrich Chemical) was prepared in dimethyl sulfoxide (DMSO, <0.5% in a final culture concentration) for cell culture.
8
Pancreatic Cancer Cell Lines Migration
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Two human pancreatic cancer cell lines, AsPC-1 (CRL-1682™) and BxPC-3 (CRL-1687™), were obtained from ATCC (Manassas, VA, USA). Cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (Gibco, Grand Island, NY, USA), 100 μg/mL of streptomycin, 100 IU/mL of penicillin, and 2 mM L-glutamine. Cells were placed in the humidified incubator with 5% CO2 at 37 °C. Human epidermal growth factor (EGF) was purchased from Sigma-Aldrich (St. Louis, MO, USA). AsPC-1 cells with or without CD44V3 were treated by recombinant EGF (50 nmol/L) for 24 h, then transwell migration and invasion assays were performed.
9
Breast Cancer Cell Line Cultivation and Signaling Pathway Analysis
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Human breast cancer MDA-MB-231 and MCF-7 cells were obtained from American Type Culture Collection (Manassas, VA, USA) and maintained in Dulbecco's modified Eagle's medium (HyClone; GE Healthcare, Chicago, IL, USA) with 10% fetal bovine serum (HyClone; GE Healthcare). Cells were incubated in a humidified atmosphere at 37°C with 5% CO2 (22 (link)). Human epidermal growth factor (EGF) and human transforming growth factor-β1 (TGF-β1) were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). The isolation of eupatolide from I. britannica and its chemical structure have been reported previously (23 (link)). Antibodies against Snail (cat. no. ab53519), Slug (cat. no. ab27568), SMAD4 (cat. no. ab40759) and phospho-EGFR (cat. no. ab76195) were purchased from Abcam (Cambridge, UK). Antibodies against vimentin (cat. no. 3932), phospho-AKT (Ser473; cat. no. 9271), ERK1/2 (cat. no. 9102), phospho-ERK1/2 (cat. no. 9106), EGF receptor (EGFR; cat. no. 4267), phospho-SMAD3 (Ser423/425; cat no. 9520), SMAD3 (cat no. 9523), phospho-SMAD1/5 (cat. no. 9516), SMAD1 (cat. no. 6944) and SMAD5 (cat. no. 12534) were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA). The antibody specific for E-cadherin was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).
10
Cell Culture Reagents and Supplies
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The
following reagents were obtained from
commercial sources as indicated: Dulbecco’s Modified Eagle’s
Medium (DMEM)/high glucose containing 4.5 g/L glucose and 4.0 mMl -glutamine (HyClone); fetal bovine serum (FBS), heat-inactivated
(Omega Scientific);l -glutamine, 200 mM (Gibco); penicillin/streptomycin
solution 50× (Mediatech); DMEM/Ham’s Nutrient Mixture
F12 containing 2.5 mMl -glutamine, 3151 mg/L dextrose, and
55 mg/L sodium pyruvate (Sigma-Aldrich); horse serum (Sigma-Aldrich);
50 μM hydrocortisone solution (Sigma-Aldrich); human insulin
solution (Sigma-Aldrich); cholera toxin (Sigma-Aldrich); human Epidermal
Growth Factor (EGF), recombinant (Sigma-Aldrich); 0.25% Trypsin-EDTA
(Gibco); nuclease-free sterile water (Fisher Scientific); molecular
biology grade DMSO (Sigma-Aldrich); ICI 182,780 (faslodex) (Tocris
Bioscience).
following reagents were obtained from
commercial sources as indicated: Dulbecco’s Modified Eagle’s
Medium (DMEM)/high glucose containing 4.5 g/L glucose and 4.0 mM
(Omega Scientific);
solution 50× (Mediatech); DMEM/Ham’s Nutrient Mixture
F12 containing 2.5 mM
55 mg/L sodium pyruvate (Sigma-Aldrich); horse serum (Sigma-Aldrich);
50 μM hydrocortisone solution (Sigma-Aldrich); human insulin
solution (Sigma-Aldrich); cholera toxin (Sigma-Aldrich); human Epidermal
Growth Factor (EGF), recombinant (Sigma-Aldrich); 0.25% Trypsin-EDTA
(Gibco); nuclease-free sterile water (Fisher Scientific); molecular
biology grade DMSO (Sigma-Aldrich); ICI 182,780 (faslodex) (Tocris
Bioscience).
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