Hb egf

Manufactured by R&D Systems

Sourced in United States

HB-EGF is a recombinant human heparin-binding epidermal growth factor-like growth factor. It is a member of the epidermal growth factor family and plays a role in cell proliferation, differentiation, and migration.

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Lab products found in correlation

Epidermal growth factor (egf), by R&D Systems (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Il 33, by R&D Systems (2 mentions) Leukemia inhibitory factor, by Thermo Fisher Scientific (2 mentions) Recombinant egf, by R&D Systems (2 mentions) Doxycycline, by Takara Bio (2 mentions) Plx4720, by Selleck Chemicals (2 mentions) Alamarblue cell viability reagent, by Thermo Fisher Scientific (2 mentions) Human phospho rtk array kit, by R&D Systems (2 mentions) α tubulin, by Cell Signaling Technology (2 mentions) P rtk, by R&D Systems (2 mentions) Sirna against mitf and non targeting control sirna, by Horizon Discovery (2 mentions) Azd6244, by Selleck Chemicals (2 mentions) Celltiter glo cell viability assay, by Promega (2 mentions) U0126, by Selleck Chemicals (2 mentions) Vemurafenib, by Selleck Chemicals (2 mentions) P akt, by Cell Signaling Technology (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) P erk, by Cell Signaling Technology (2 mentions) Fibroblast growth factor 2 (fgf2), by R&D Systems (2 mentions)

34 protocols using hb egf

Cytokine Induction in Immune Cells

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To determine the induction of cytokines expression, FACS-sorted T cells and mLN cells or splenocytes were cultured in 96-well plates in IMDM medium, 10% FCS, 1% l-glutamine, 1% penicillin/streptomycin (GIBCO), 5 × 10⁻⁵ M 2-mercaptoethanol in the presence or absence of Monoensin (Invitrogen) (10 μM) and rm IL-33 (PeproTech) (10 ng/mL) or anti-CD3 (BD, 145-2C11) (2 μg/mL) for six hours. Alternatively, intracellular amounts of p-EGFR (Y1068 and Y992), p-ERK and IκBα were analyzed after 30 minutes of stimulation with IL-33 (10 ng/mL), anti-CD3 (2 μg/mL), AREG (R&D) (10 ng/mL) or HB-EGF (R&D) (10 ng/mL). To analyze EGFR or AREG induction, splenocytes or FACS-sorted T cells were re-stimulated overnight with H. polygyrus excretory-secretory products (HES; Johnston et al., 2015 ) (1 μg/mL), adult worm extract (HEX) (10 μg/mL), anti-CD3 (2 μg/mL), IL-7 (PeproTech) (20 ng/mL), IL-2 (BD) (1 ng/mL), TSLP (eBiosciences) (20 ng/mL) and IL-33 (10 ng/mL). Inhibitors were used at the following concentrations: MEK inhibitor (Promega)(10 μM), Gefitinib (LC laboratories) (1 μM) and Marimastat (AbCam) (25 μM). The inhibitory effect of Marimastat (25 μM) was checked by its ability to prevent the release of activated Amphiregulin by transfected HEK cells. Under these conditions cell viability was higher than 90% as compared to 97% at the start of the experiment.

Minutti C.M., Drube S., Blair N., Schwartz C., McCrae J.C., McKenzie A.N., Kamradt T., Mokry M., Coffer P.J., Sibilia M., Sijts A.J., Fallon P.G., Maizels R.M, & Zaiss D.M. (2017). Epidermal Growth Factor Receptor Expression Licenses Type-2 Helper T Cells to Function in a T Cell Receptor-Independent Fashion. Immunity, 47(4), 710-722.e6.

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CD44+ Cell Sorting for Astrocyte Induction

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Approximately 4–5×10⁶ cells were resuspended in 100μl of sort buffer (2% FBS, 1mM EDTA in PBS) and CD44 conjugated with Alexa 647 was incubated as per manufacturer’s instructions on ice for 20–30 minutes. Cells were washed 2 times with sort buffer and taken to the sorter. CD44 positive cells were maintained in astrocyte induction media (N2 media with 10ng/ml HB-EGF (R&D Systems) and 10ng/ml Leukemia inhibitory factor (Peprotech) without doxycycline.

Tchieu J., Calder E.L., Guttikonda S.R., Gutzwiller E.M., Aromolaran K.A., Steinbeck J.A., Goldstein P.A, & Studer L. (2019). NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells. Nature biotechnology, 37(3), 267-275.

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Astrocyte Derivation from CD44+ Cells

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were maintained on PO/Lam/FN coated dishes in astrocyte media consisting of N2 media with 10ng/ml of HB-EGF (R&D Systems, 259-HE). After sorting, the CD44 positive cells were passaged every week for 4 weeks and then passaged every other week or until the astrocyte processes started to detach.

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Ex Vivo Salivary Gland Epithelial Culture

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In all ex vivo epithelial rudiment culture experiments, SG were derived from CD1 timed pregnant mice at E13. E13 SG epithelia were freshly isolated as previously described^{11 (link),12}. Briefly, the SG epithelium was separated from the mesenchyme using dispase II (Gibco) treatment followed by mechanical dissection and cultured in 15μl of laminin-1 (R&D Systems) on a nucleopore filter floating above 200 μL of serum-free media (DMEM/F12; Thermo Fisher Scientific) containing penicillin-streptomycin (Gibco), 50μg/ml holo-transferrin (HT), 150μg/ml ascorbic acid (AsA) and 200ng/ml human recombinant NRG1 isoform SMDF (R&D Systems), or 20ng/ml HB-EGF (R&D Systems) for up to 120 h. For inhibitor experiments, epithelia were cultured at 37°C in a humidified 5% CO₂/95% air atmosphere in complete media in the presence of 0.2% DMSO (Sigma-Alrdrich), 5μM of mTORC1 inhibitor rapamycin (Biotang Inc.), or 1μM mTORC1/2 inhibitor Torin-1 (Cell Signaling Technologies). In all experiments, media was refreshed daily.

May A.J., Mattingly A.J., Gaylord E.A., Griffin N., Sudiwala S., Cruz-Pacheco N., Emmerson E., Mohabbat S., Nathan S., Sinada H., Lombaert I.M, & Knox S.M. (2022). Neuronal-epithelial cross-talk drives acinar specification via NRG1-ERBB3-mTORC2 signaling. Developmental cell, 57(22), 2550-2565.e5.

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Quantifying Growth Factors in Stool

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Frozen stool specimens were resuspended in PBS, homogenized, and analyzed by enzyme-linked immunosorbent assay (ELISA) for human EGF (R&D Systems), human amphiregulin (AREG) (R&D Systems), human TGF-α (R&D Systems), and human heparin-binding EGF (HB-EGF) (R&D Systems), per the manufacturer’s protocol. For mice, stool was collected in PBS, homogenized, and analyzed by ELISA for murine EGF (R&D Systems), per manufacturer’s protocol.

Knoop K.A., Coughlin P.E., Floyd A.N., Ndao I.M., Hall-Moore C., Shaikh N., Gasparrini A.J., Rusconi B., Escobedo M., Good M., Warner B.B., Tarr P.I, & Newberry R.D. (2020). Maternal activation of the EGFR prevents translocation of gut-residing pathogenic Escherichia coli in a model of late-onset neonatal sepsis. Proceedings of the National Academy of Sciences of the United States of America, 117(14), 7941-7949.

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Biomarker Profiling for Metabolic Health

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Levels of HB-EGF (R&D Systems, USA), IL-17A (R&D Systems) and MPO (R&D Systems) were measured by ELISA, of which the reference ranges were 0–35 ng/L, 0–20 pg/mL and 0–3.4 µg/L, respectively. Blood routine examination was performed using a blood cell analysis workstation (XE-AlphaN, Sysmex Corp., Kobe, Japan).
Total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), serum uric acid, Cr, blood urea nitrogen (BUN) and triglycerides (TG) were measured using an automatic biochemical analyzer (Aeroset, Abbott Laboratories, Abbott Park, Illinois, USA).

Jiang H., Zhang H., Yang Y, & Yang X. (2020). Associations of myeloperoxidase, interleukin-17A and heparin-binding EGF-like growth factor levels with in-stent restenosis after percutaneous coronary intervention: a single-centre case–control study in China. BMJ Open, 10(11), e039405.

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Signaling Pathway Protein Profiling

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Equal amounts of protein were separated by electrophoresis through NuPage Bis-Tris 4-12% gradient gels (Invitrogen), proteins were transferred onto nitrocellulose pore membranes using the iBlot system and protocol from Invitrogen (Carlsbad, CA). PTEN, pAKT (T308), pAkt (Ser473), pERK (T202/Y204), Total EGFR, pEGFR (Y1068), and pS6 (S235/236) antibodies were obtained from Cell Signaling Technology (Danvers, MA). pRSK (T359/S363) antibody was obtained from EMD Millipore (Billerica, Massachusetts). ELISA kits for AREG, βcellulin, HRG-1, EGF, HB-EGF, and TGFα, were obtained from R&D systems (Minneapolis, MN) and carried out according to provided instructions.

Edgar K.A., Crocker L., Cheng E., Wagle M.C., Wongchenko M., Yan Y., Wilson T.R., Dompe N., Neve R.M., Belvin M., Sampath D., Friedman L.S, & Wallin J.J. (2014). Amphiregulin and PTEN evoke a multimodal mechanism of acquired resistance to PI3K inhibition. Genes & Cancer, 5(3-4), 113-126.

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Astrocyte Derivation from CD44+ Cells

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Isolation and Characterization of Murine Adipose-Derived Stromal Cells

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Murine ASC were obtained from 6–8 weeks old C57Bl/6J mice (Charles River Laboratories). The isolation of stromal-vascular fraction from adipose tissue was carried out as previously described^{27 (link),38 (link)}. Briefly, extracellular matrix was digested with Collagenase A (Roche) and centrifuged at 1200 g. The pellet obtained was re-suspended and the stromal fraction was collected by sequential centrifugation and filtration steps. Cells were then cultured in complete medium (DMEM + glucose + GlutaMAX ITM + 15% heat-inactivated adult bovine serum + penicillin/streptomycin; all from Invitrogen) supplemented with 50 ng/ml HB-EGF (R&D Systems). The immune phenotype of murine ASC was characterized by using monoclonal antibodies (mAb) specific for CD106, CD9, CD44, CD80, CD138, and Stem cells antigen 1 (Sca1). In addition, the absence of hematopoietic (CD45, CD11c, and CD34) and endothelial markers (CD31) was assessed as previously described^{6 (link)}. All mAbs were purchased from BD Pharmingen (San Diego). Isotype-matched antibodies were used as controls. For immunophenotypic analysis, ASC were incubated at 4 °C for 10 min with 15% adult bovine serum followed by incubation with the specific mAb at 4 °C for 30 min. At least 10,000 events were analyzed by flow cytometry on a FACScalibur (Becton Dickinson) using the Cell Quest software (Becton Dickinson).

Farinazzo A., Angiari S., Turano E., Bistaffa E., Dusi S., Ruggieri S., Bonafede R., Mariotti R., Constantin G, & Bonetti B. (2018). Nanovesicles from adipose-derived mesenchymal stem cells inhibit T lymphocyte trafficking and ameliorate chronic experimental autoimmune encephalomyelitis. Scientific Reports, 8, 7473.

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Cell Signaling Pathway Inhibitors Study

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Alectinib, crizotinib, and 17-DMAG were purchased from Seleck Chemicals(Houston, TX). Recombinant EGF, TGF-α, and HB-EGF were purchased from R&D Systems(Minneapolis, MN). Recombinant HGF was prepared as described in a previous study [38 (link)].

Tanimoto A., Yamada T., Nanjo S., Takeuchi S., Ebi H., Kita K., Matsumoto K, & Yano S. (2014). Receptor ligand-triggered resistance to alectinib and its circumvention by Hsp90 inhibition in EML4-ALK lung cancer cells. Oncotarget, 5(13), 4920-4928.

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