Recombinant human egf af 100 15

Manufactured by Thermo Fisher Scientific

Sourced in United States

Recombinant human EGF (AF-100-15) is a laboratory reagent produced by Thermo Fisher Scientific. It is a purified, recombinant form of the human epidermal growth factor (EGF) protein. EGF is a signaling molecule that plays a role in cell growth and proliferation. The recombinant human EGF product can be used in various in vitro cell culture and research applications.

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Recombinant human EGF (229 citations) Human EGF (156 citations) AF-100-15 (122 citations) Recombinant EGF (35 citations) EGF (AF-100–15, (17 citations) Human EGF (AF-100-15, (5 citations) AF-100-15-100 (4 citations)

7 protocols using recombinant human egf af 100 15

Investigating Receptor Signaling Pathways

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The following antibodies were used according to the protocols supplied by the manufacturers: anti-pERK1/2 (#9101, Cell Signaling Technology, Danvers, MA, USA), anti-ERK1/2 (#4695, Cell Signaling Technology), anti-GAPDH (ENM0040, Elabscience Biotechnology Inc., Houston, TX, USA), anti-pEGFR (12A3) (sc-57542, Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-EGFR (C-2) (sc-377229, Santa Cruz Biotechnology Inc.), anti-pAKT (#9271, Cell Signaling Technology), anti-AKT (#9272, Cell Signaling Technology), anti-Cyclin D1 (sc-753, Santa Cruz Biotechnology Inc.), anti-p21 (sc-397, Santa Cruz Biotechnology Inc.), and anti-beta Tubulin (Sigma), anti-alpha actin (Sigma). All the secondary antibodies (HRP-conjugated anti-rabbit and anti-mouse) were purchased from Santa Cruz Biotechnology Inc.
Drug formulations: gefitinib (ZD1839) and MG132 (S2619) were purchased from Selleckchem; AZD9291 was obtained from AstraZeneca; BAPTA_AM (HB0981) was from HelloBio.
Recombinant Human EGF (AF-100-15) was from PeproTech (London, UK).

Modica T.M., Dituri F., Mancarella S., Pisano C., Fabregat I, & Giannelli G. (2019). Calcium Regulates HCC Proliferation as well as EGFR Recycling/Degradation and Could Be a New Therapeutic Target in HCC. Cancers, 11(10), 1588.

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Signaling Pathway Protein Analysis

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Antibodies to pAKT (cat# 4060), total AKT (cat# 9272), pEGFR(Y845) (cat #2231), pEGFR(Y1092), pEGFR (Y1068) (cat# 3777), pEGFR (Y1173), total EGFR (cat # 4267; # 2232), Her2/ErbB2, c-Met, pFAK, FAK, pErk1/2 (42/44) (cat # 9101; cat # 4370), Erk1/2 (42/44) (cat # 4695) and GAPDH (all from Cell Signaling), β-adaptin (BD Biosciences, cat # 610382), EEA1 (Ab70521), and Actin were from Abcam. total EGFR antibody (Sc-03) was from SantaCruz Biotechnology. The HRP-conjugated secondary antibodies were from Jackson ImmunoResearch. Recombinant human EGF (AF-100-15) was obtained from PeproTech, custom BUB1 siRNA, as well as non-silencing siRNA (NSS) were obtained from GE-Dharmacon. SD208 (Tocris), Erlotinib (gift from Genentech), Cetuximab (Bristol Myer Squibb), 2OH-BNPP1 was synthesized in-house as described by Jiang et al., [40 ].
Cycloheximide was obtained from Acros Organics/Fischer Scientific, while DSS was from ThermoFisher Scientific. FuGENE 6 was from Roche while Lipofectamine 2000 was from Invitrogen/ThermoFisher Scientific. ProLong Gold anti-fade mounting media (without DAPI) was purchased from Invitrogen/ThermoFisher Scientific.

Nyati S., Young G., Speers C., Nyati M.K, & Rehemtulla A. (2023). Budding uninhibited by benzimidazoles-1 (BUB1) regulates EGFR signaling by reducing EGFR internalization. Aging (Albany NY), 15(13), 6011-6030.

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Sphere Formation of HCC Cells

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5 × 10⁴ HCC cells were placed in ultra-low binding 6-well dishes and grown in DMEM/F12 medium. Animal-Free Recombinant Human EGF (AF-100-15, Peprotech), Recombinant Human FGF-basic (100-18B, Peprotech) and N-2 supplement (17502-048, ThermoFisher) were added to the medium to ensure cell sphere formation. When the spheres formed by the cells grew to 8 days, they were viewed and photographed using a microscope.

Wu Q., Qiu Y., Guo J., Yuan Z., Yang Y., Zhu Q., Zhang Z., Guo J., Wu Y., Zhang J., Huang D., Tu K, & Hu X. (2024). USP40 promotes hepatocellular carcinoma cell proliferation, migration and stemness by deubiquitinating and stabilizing Claudin1. Biology Direct, 19, 13.

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Regulation of Luciferase Activity by EGF, P38, and miR-675

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Cells cultured on 96‐well plates were transiently co‐transfected with firefly luciferase reporter plasmids (0.2 μg) and ranilla luciferase co‐reporter plasmids (pRL‐TK, 0.02 μg). Then, these cells were simultaneously or subsequently treated with EGF (20 ng/mL), P38 MAPK inhibitor SB203580 (10 μmol/L), or miR‐675‐5p mimics (20 μmol/L, add 0.4 μL into 100 μL medium per well) for the indicated time periods. Next, the cellular luciferase activity was measured using Dual‐luciferase^® Reporter Assay System kit (#E1910, Promega) according to the manufacturer's instructions. Results were calculated as the ratios between the activity of firefly luciferase and ranilla luciferase. The recombinant human EGF (#AF‐100‐15) was purchased from PeproTech (Rocky Hill, NJ, USA), and P38 MAPK inhibitor SB203580 (#HY‐10256) was purchased from MedChem Express (Monmouth Junction, NJ, USA).

Liu Z., Ning F., Cai Y., Sheng H., Zheng R., Yin X., Lu Z., Su L., Chen X., Zeng C., Wang H, & Liu L. (2021). The EGFR‐P38 MAPK axis up‐regulates PD‐L1 through miR‐675‐5p and down‐regulates HLA‐ABC via hexokinase‐2 in hepatocellular carcinoma cells. Cancer Communications, 41(1), 62-78.

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Molecular Analysis of 6-Gingerol Effects

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Roswell Park Memorial Institute-1640 (RPMI-1640) medium, penicillin-streptomycin solution, and trypsin-EDTA (0.05%) were all purchased from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA). 6-Gingerol (cat no. 23513-14-6) was purchased from TCI (Tokyo Chemical Industry Co., Tokyo, Japan). Fetal bovine serum (FBS) was obtained from Sigma–Aldrich (Merck KGaA, St. Louis, MO, USA). Recombinant human EGF (AF-100-15) was purchased from Peprotech, Inc. (Rocky Hill, NJ, USA). Antibodies specific for CDK4 (sc-23896), BCL-2 (sc-7382), p21 (sc-6246), and cyclin E (sc-377100) together with secondary antibodies (anti-mouse (sc-516102) and anti-rabbit (sc-2357)) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies specific for pEGFR (#3777), EGFR (#4267), pSTAT3 (#9145), STAT3 (#9139), cyclin D1 (#2922), p27 (#3686), p53 (#9282), pCHK1 (#2348), pATM (#5883), pBRCA1 (#9009), pHistone H2A.X (#9718), pCHK2 (#2197), BAX (#2772), Casp 9 (#9502), C-Casp 9 (#9505), cytochrome c (#11940), BCL-xL (#2764), pMDM2 (#3521), MDM2 (#86934), pSrc (#6943), Src (#2109), and GAPDH (#2118) were obtained from Cell Signaling Technology (Beverly, MA, USA). Finally, iNOS antibody (NB300-650) was purchased from Novus Biologicals (Littleton, CO, USA).

Sp N., Kang D.Y., Lee J.M., Bae S.W, & Jang K.J. (2021). Potential Antitumor Effects of 6-Gingerol in p53-Dependent Mitochondrial Apoptosis and Inhibition of Tumor Sphere Formation in Breast Cancer Cells. International Journal of Molecular Sciences, 22(9), 4660.

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Molecular Probes for Cell Signaling

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Raphin1 acetate (HY-123960A), Sanguinarine chloride (HY-N0052A), KY-226 (HY-120327), SHP099 (HY-100388), NSC95397 (HY-108543), and BCI (HY-115502) were purchased from MedChemExpress. BIRB796 (Axon 1358) was purchased from Axon Medchem. Recombinant human EGF (AF-100-15) was purchased from Peprotech.

Eguchi A, & Olsen J.V. (2024). Phosphoproteomic investigation of targets of protein phosphatases in EGFR signaling. Scientific Reports, 14, 7908.

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EGF-Induced EGFR and PI3K Signaling

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Whole-cell lysates were prepared by radioimmunoprecipitation assay buffer (EMD Millipore, Burlington, MA, USA) containing phosphatase and protease inhibitors. Antibodies specific for β-actin (sc-47778), p53 (sc-126) and secondary antibodies (antimouse (sc-516102), and antirabbit (sc-2357) antibody) were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). pEGFR (#2234), EGFR (#3776s), pAKT (#4060), AKT (#4691), pPI3K (#4228), and PI3K (#4257) antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). PD-L1 (R30949) antibody was purchased from NSJ Bioreagents (San Diego, CA, USA). Recombinant human EGF (AF-100-15) was purchased from PeproTech Inc. (Rocky Hill, NJ, USA).

Kang D.Y., Sp N., Jo E.S., Rugamba A., Hong D.Y., Lee H.G., Yoo J.S., Liu Q., Jang K.J, & Yang Y.M. (2020). The Inhibitory Mechanisms of Tumor PD-L1 Expression by Natural Bioactive Gallic Acid in Non-Small-Cell Lung Cancer (NSCLC) Cells. Cancers, 12(3), 727.

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