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Recombinant human egf

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Recombinant human EGF is a protein produced in E. coli cells. It is a member of the epidermal growth factor family and plays a role in cell growth and differentiation.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions) N2 supplement, by Thermo Fisher Scientific (6 mentions) Glutamax, by Thermo Fisher Scientific (6 mentions) Hydrocortisone, by Merck Group (5 mentions) Heparin, by Merck Group (5 mentions) L glutamine, by Corning (5 mentions) Recombinant human bfgf, by R&D Systems (5 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions) Cholera toxin, by Merck Group (4 mentions) Human recombinant fgf2, by Thermo Fisher Scientific (4 mentions) Neurobasal medium, by Thermo Fisher Scientific (4 mentions) B27 supplement, by Thermo Fisher Scientific (4 mentions) Recombinant human wnt3a, by R&D Systems (4 mentions) Anti akt, by Cell Signaling Technology (4 mentions) β actin, by Merck Group (4 mentions) Bovine insulin, by Merck Group (3 mentions) Horse serum, by Thermo Fisher Scientific (3 mentions) Accutase, by Innovative Cell Technologies (3 mentions) Basic fibroblast growth factor (bfgf), by R&D Systems (3 mentions) Dmem f12, by Thermo Fisher Scientific (3 mentions)

Spelling variants of this product

Human EGF (24 citations) Recombinant EGF (15 citations) Human recombinant bFGF and EGF (8 citations) Recombinant human EGF protein (8 citations) Recombinant human HB-EGF (6 citations) Human recombinant epidermal growth factor (EGF, (3 citations) Recombinant Human EGF Protein (236‐EG) (2 citations) Recombinant human EGF and FGF (2 citations)

71 protocols using recombinant human egf

1

Maintenance of Patient-Derived Glioblastoma Stem Cells

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Patient-derived GSC MGG8 was obtained from Massachusetts General Hospital as previously described (34 (link)). Cells were maintained in the stem cell culture medium [Neurobasal medium (Gibco; Invitrogen), supplemented with 3 mM of L-glutamine (Cellgro), B27 supplement (Gibco; Invitrogen), N2 supplement (Gibco; Invitrogen), heparin (Sigma), penicillin/streptomycin/amphotericin B (Cellgro), human recombinant FGF-2 (Peprotech) for final 20 ng/ml, human recombinant EGF (R and D systems) for final 20 ng/ml]. Furthermore, cells were cultured in an atmosphere containing 5% CO2 at 37°C. Recombinant human bone BMP-4 was purchased from R&D Systems.
Koguchi M., Nakahara Y., Ito H., Wakamiya T., Yoshioka F., Ogata A., Inoue K., Masuoka J., Izumi H, & Abe T. (2019). BMP4 induces asymmetric cell division in human glioma stem-like cells. Oncology Letters, 19(2), 1247-1254.
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2

Kinetics of EGF-induced RAS activation

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5 X 106 cells were plated in 10 cm2 plates and serum starved overnight. Cells were treated with 50 ng/mL human recombinant EGF (R&D Systems). Protein lysates were harvested 0, 5, 15, 30 and 60 minutes after EGF stimulation. Cells were washed twice with cold PBS and lysed with Mg2+ lysis buffer supplemented by protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (Calbiochem). The RAS-GTP assay was carried out using the RAS Activation Assay kit (Millipore) according to manufacturer’s instructions. Activated RAS was precipitated with purified GST-RAF-RBD agarose beads by pre-incubating 1 mg of whole cell lysates with GST-RAF-RBD pre-bound to gluthiathione-sepharose. Bound RAS was subjected to SDS-PAGE electrophoresis and immunoblotting analysis using anti-KRAS antibody (Millipore).
Wong G.S., Zhou J., Liu J.B., Wu Z., Xu X., Li T., Xu D., Schumacher S.E., Puschhof J., McFarland J., Zou C., Dulak A., Henderson L., Xu P., O’Day E., Rendak R., Liao W.L., Cecchi F., Hembrough T., Schwartz S., Szeto C., Rustgi A.K., Wong K.K., Diehl J.A., Jensen K., Graziano F., Ruzzo A., Fereshetian S., Mertins P., Carr S.A., Beroukhim R., Nakamura K., Oki E., Watanabe M., Baba H., Imamura Y., Catenacci D, & Bass A.J. (2018). Targeting wild-type KRAS amplified gastroesophageal cancer through combined MEK and SHP2. Nature medicine, 24(7), 968-977.
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3

Antibody and Reagent Usage in Study

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The following primary antibodies and reagents were used in this study: Antibodies against phospho-AKT(Ser-473), AKT, Caspase 8, cleaved Caspase 3, EGFR, cleaved PARP, phospho-S6 ribosomal protein(Ser-235/236), S6 ribosomal protein, phospho-p-44/42MAPK(ERK1/2) (Thr202/Tyr204), and p-44/42MAPK(ERK1/2) (Cell Signaling); anti-phospho-EGFR(Tyr-1068) (Abcam); anti-α Tubulin (Sigma); anti-DR5 (Santa Cruz); Cetuximab (ImClone Systems); Erlotinib (Selleck Chemicals); human recombinant EGF (R&D Systems); anti-human CD261 (DR4)PE and anti-human CD262 (DR5)PE(eBioscience).
Zhu Y., Bassoff N., Reinshagen C., Bhere D., Nowicki M.O., Lawler S.E., Roux J, & Shah K. (2017). Bi-specific molecule against EGFR and death receptors simultaneously targets proliferation and death pathways in tumors. Scientific Reports, 7, 2602.
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4

Cell Line Maintenance and Characterization

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HEK 293 and PlatE cells were maintained in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% FCS (Serana). MCF-10A SgK223 KO cells were as previously described10 (link) and were maintained in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (Invitrogen) supplemented with 5% (v/v) horse serum (Invitrogen), 20 ng ml−1 human recombinant EGF (R&D Systems), 0.5 μg ml−1 hydrocortisone (Sigma), 100 ng ml−1 cholera toxin (Sigma), 10 μg ml−1 bovine insulin (Sigma), 50 units per ml penicillin G (Invitrogen) and 50 μg ml−1 streptomycin sulfate (Invitrogen). Retroviral infection of these cells, transfection of HEK293 cells, and co-immunoprecipitation studies, were also undertaken as previously reported10 (link). Western blot analysis were performed using the following antibody dilutions: Flag (1:1,000), HA (1:1,000), Actin (1:5,000), SgK223 (1:500), SgK269 (1:500) (Supplementary Table 1). Raw uncropped western blot scans are in Supplementary Figs. 4 and 7.
Patel O., Griffin M.D., Panjikar S., Dai W., Ma X., Chan H., Zheng C., Kropp A., Murphy J.M., Daly R.J, & Lucet I.S. (2017). Structure of SgK223 pseudokinase reveals novel mechanisms of homotypic and heterotypic association. Nature Communications, 8, 1157.
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5

Isolation and Culture of Primary Glioma Stem Cells

Cited 1 time
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Primary glioma stem cells (GSCs) were derived from resected tumor tissue of GBM patients undergoing treatment at the Massachusetts Generals Hospital (provided by Dr. Hiroaki Wakimoto) or The Ohio State University James Comprehensive Cancer Center, in accordance with the appropriate institutional review board approval. All GSC lines used in this study including proneural cells (BT107, MGG6, MGG8 and PN157) as well as mesenchymal cells (MES83 and MES326) were previously characterized.[19 (link),22 (link),39 (link)–42 ] Cells were cultured as neurospheres and maintained in Gibco Dulbecco’s Modified Eagle Medium: Nutrient Mixture F-12 (Thermo Fisher Scientific) containing the following supplements: 3 mm L-glutamine, 1:50 B27 (Life Technologies), 20 ng/mL human recombinant EGF (R & D Systems), and 20 ng/mL human recombinant bFGF-2 (Peprotech). Nanoparticle tracking analysis was performed to confirm that the cell culture medium used had no detectable amounts of EVs. Neurospheres were dissociated using Accutase (Innovative Cell Technologies) before every passage and cultured in a humidified incubator with 5% CO2 at 37 °C. Cells were counted using a Bright-Line™ Hemocytometer (Sigma).
Schweiger M.W., Li M., Giovanazzi A., Fleming R.L., Tabet E.I., Nakano I., Würdinger T., Chiocca E.A., Tian T, & Tannous B.A. (2020). Extracellular Vesicles Induce Mesenchymal Transition and Therapeutic Resistance in Glioblastomas through NF-κB/STAT3 Signaling. Advanced biosystems, 4(12), e1900312.
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6

Cell Culture Conditions for Cell Lines

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MCF-10A cells obtained from the ATCC (USA) were cultured in Dulbecco’s Modified Eagle Medium /nutrient mixture F-12 (Invitrogen) with 5% (v/v) horse serum (Invitrogen), 0.5% μg/ml hydrocortisone (Sigma), 10 μg/ml bovine insulin (Sigma), 100 ng/ml cholera toxin (Sigma) and 20 ng/ml human recombinant EGF (R&D Systems). HeLa, MDA-MB-231 and MDA-MB-468 cells were obtained from ATCC (USA) and maintained in RPMI 1640 media (Gibco) with 10% (v/v) Fetal Bovin Serum, 10 μg/ml Insulin and 20 mM HEPES. All cells grew in incubators with a humidified atmosphere containing 5% CO2 and 95% air at 37°C.
Ghomlaghi M., Theocharous M., Hoang N., Shin S.Y., von Kriegsheim A., O’ Neill E., Zhang T, & Nguyen L.K. (2024). Integrative modeling and analysis of signaling crosstalk reveal molecular switches coordinating Yes-associated protein transcriptional activities. iScience, 27(3), 109031.
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7

Angiogenesis Modulation in Matrigel Plugs

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Male C57B/l6 mice were injected subcutaneously above the pelvis area with 200 μl of liquid growth factor-reduced Matrigel combined either with EG-VEGF (Miltenyi Biotechnologies, Paris, France) or with human recombinant EGF (R&D Systems, Metz, France) (500 ng/ml, each), UT agonists (UII, mUII, URP, and UII4–11) at 50 ng/ml, UT antagonists/biased ligand palosuran or urantide at 1 μg/ml, or alone as a negative control. Subcutaneous plug incubation continued for 21 days or until mice had to be sacrificed following institutional ethical guidelines criteria. Matrigel plugs were resected and immediately frozen in a −40°C isopentane solution until immunohistochemistry.
Le Joncour V., Guichet P.O., Dembélé K.P., Mutel A., Campisi D., Perzo N., Desrues L., Modzelewski R., Couraud P.O., Honnorat J., Ferracci F.X., Marguet F., Laquerrière A., Vera P., Bohn P., Langlois O., Morin F., Gandolfo P, & Castel H. (2021). Targeting the Urotensin II/UT G Protein-Coupled Receptor to Counteract Angiogenesis and Mesenchymal Hypoxia/Necrosis in Glioblastoma. Frontiers in Cell and Developmental Biology, 9, 652544.
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8

Intrahepatic Cholangiocarcinoma Cell Lines

Cited 2 times
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The human intrahepatic CCA cell lines CCLP1, HUCCT1 (a kind gift from A.J. Demetris, Pittsburgh) and CCA4 were previously described6 (link). The human immortalized non-malignant cholangiocyte cell line H69 was kindly provided by Dr. G. J. Gores (Mayo Clinic, Rochester). The cell lines were grown at 37 °C in a humidified incubator at 5% CO2 as monolayers (MON) in DMEM medium or spheres (SPH) in anchoring-independent conditions with selective serum-free DMEM/F12 medium supplemented with 1X B27 supplement minus vitamin A (Life Technologies, Monza, Italy), human recombinant EGF (R&D System, Milano, Italy) (20 ng/mL) and bFGF (R&D System) (20 ng/mL)4 (link),6 (link). Cells were treated for 18 hours with 100 µM Desferioxamine (DFO) (Sigma Aldrich, Milano, Italy) or 100 µg/ml ferric ammonium citrate (FAC) (Sigma Aldrich) or increasing concentrations of erastin (Sigma Aldrich).
Raggi C., Gammella E., Correnti M., Buratti P., Forti E., Andersen J.B., Alpini G., Glaser S., Alvaro D., Invernizzi P., Cairo G, & Recalcati S. (2017). Dysregulation of Iron Metabolism in Cholangiocarcinoma Stem-like Cells. Scientific Reports, 7, 17667.
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9

Kinetics of EGF-induced RAS activation

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5 X 106 cells were plated in 10 cm2 plates and serum starved overnight. Cells were treated with 50 ng/mL human recombinant EGF (R&D Systems). Protein lysates were harvested 0, 5, 15, 30 and 60 minutes after EGF stimulation. Cells were washed twice with cold PBS and lysed with Mg2+ lysis buffer supplemented by protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (Calbiochem). The RAS-GTP assay was carried out using the RAS Activation Assay kit (Millipore) according to manufacturer’s instructions. Activated RAS was precipitated with purified GST-RAF-RBD agarose beads by pre-incubating 1 mg of whole cell lysates with GST-RAF-RBD pre-bound to gluthiathione-sepharose. Bound RAS was subjected to SDS-PAGE electrophoresis and immunoblotting analysis using anti-KRAS antibody (Millipore).
Wong G.S., Zhou J., Liu J.B., Wu Z., Xu X., Li T., Xu D., Schumacher S.E., Puschhof J., McFarland J., Zou C., Dulak A., Henderson L., Xu P., O’Day E., Rendak R., Liao W.L., Cecchi F., Hembrough T., Schwartz S., Szeto C., Rustgi A.K., Wong K.K., Diehl J.A., Jensen K., Graziano F., Ruzzo A., Fereshetian S., Mertins P., Carr S.A., Beroukhim R., Nakamura K., Oki E., Watanabe M., Baba H., Imamura Y., Catenacci D, & Bass A.J. (2018). Targeting wild-type KRAS amplified gastroesophageal cancer through combined MEK and SHP2. Nature medicine, 24(7), 968-977.
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10

Generating Stable Cell Lines for BioID

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MCF-10A cells stably expressing the murine ecotropic receptor (MCF-10A EcoR) were used to generate stable pools of MCF-10A cells expressing NEK5-BioID (referred as to MCF-10A NEK5) or control vector (referred as MCF-10A EV) [21 (link)]. To produce stable cell lines, PlatE cells were transfected with DNA vectors using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s instructions. Viral supernatants were collected at 48 h and 72 h after transfection and filtered through 0.45 μm pore-size filter caps (Millipore). MCF-10A EcoR cells were infected with viral supernatants for 24 h in the presence of 8 μg/ml polybrene (Millipore). Successfully transduced cells were selected with puromycin (2 μg/mL) for pBABE-BioID constructs. MCF-10A cells and their derivatives were maintained in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (Invitrogen) supplemented with 5% (v/v) horse serum (Invitrogen), 20 ng/ml human recombinant EGF (R&D Systems), 0.5 μg/ml hydrocortisone (Sigma), 100 ng/ml cholera toxin (Sigma), and 10 μg/ml bovine insulin (Sigma).
de Castro Ferezin C., Lim Kam Sian T.C., Wu Y., Ma X., Chüeh A.C., Huang C., Schittenhelm R.B., Kobarg J, & Daly R.J. (2022). Identification of biological pathways and processes regulated by NEK5 in breast epithelial cells via an integrated proteomic approach. Cell Communication and Signaling : CCS, 20, 197.
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