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Recombinant hgf

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Recombinant HGF is a purified protein produced in a mammalian cell expression system. It is the human hepatocyte growth factor (HGF) protein.

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Lab products found in correlation

Cell culture insert, by Merck Group (1 mentions) Mc170 hd, by Leica (1 mentions) Recombinant fgf10, by R&D Systems (1 mentions) Dm il led microscope, by Leica (1 mentions) Recombinant tgf β1, by MyBioSource (1 mentions) Thapsigargin, by Merck Group (1 mentions) Tunicamycin, by Merck Group (1 mentions) Anti p egfr, by Cell Signaling Technology (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions) Anti p akt, by Cell Signaling Technology (1 mentions) Jnj 38877605, by Selleck Chemicals (1 mentions) Anti gab1, by Cell Signaling Technology (1 mentions) Anti p gab1, by Cell Signaling Technology (1 mentions) Pro hgf, by R&D Systems (1 mentions) Anti p erk1 2, by Cell Signaling Technology (1 mentions) Anti β actin, by Merck Group (1 mentions) Celltracker orange, by Thermo Fisher Scientific (1 mentions) Lsm 780 scope, by Zeiss (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Cell titer glo luminescence cell viability kit, by Promega (1 mentions)

Close spelling variants of this product

Recombinant human HGF (49 citations) Recombinant mouse HGF (6 citations) Recombinant human HGF protein (6 citations) Recombinant mouse HGF protein (2 citations) Murine recombinant HGF protein (2 citations)

7 protocols using recombinant hgf

1

HGF Stress Response in A549 Cells

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ER stressed A549 cells were treated with 1.6 ng/ml recombinant HGF from R&D Systems (Abingdon, UK).
Nita I., Hostettler K., Tamo L., Medová M., Bombaci G., Zhong J., Allam R., Zimmer Y., Roth M., Geiser T, & Gazdhar A. (2017). Hepatocyte growth factor secreted by bone marrow stem cell reduce ER stress and improves repair in alveolar epithelial II cells. Scientific Reports, 7, 41901.
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2

EpiSPC and rMC Co-culture Protocol

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Mono- or co-culture of EpiSPC and rMC was performed as described previously [30 ]. In brief, flow sorted cells were counted, resuspended and mixed with growth factor reduced matrix (BD Biosciences) diluted with EpiSPC medium (α-MEM, 10% FCS, 1x pen/strep, 1x insulin/transferrin/selenium, 2mM L-glutamine, 0.0002% heparin) at a 1:1 ratio. Cell suspensions were seeded in 12mm cell culture inserts (0.4μm pore size, Millipore) in a 24-well plate and incubated for 5 min at 37°C, 5% CO2 to allow gelling. EpiSPC medium was then added to the bottom wells of the plate. In selected experiments, 50ng/ml recombinant Fgf10 and 30ng/ml recombinant Hgf (both R&D systems) or anti-Fgf10 or control Ab were added to the medium at day 2 of culture. For matrix digestion a preheated dispase/collagenase I (Boehringer, Gibco) mixture (3 mg/ml) was added and a single cell suspension was obtained for re-seeding or single-cell sorting. Images were taken with a DM IL LED microscope and a corresponding camera MC170 HD (Leica).
Quantius J., Schmoldt C., Vazquez-Armendariz A.I., Becker C., El Agha E., Wilhelm J., Morty R.E., Vadász I., Mayer K., Gattenloehner S., Fink L., Matrosovich M., Li X., Seeger W., Lohmeyer J., Bellusci S, & Herold S. (2016). Influenza Virus Infects Epithelial Stem/Progenitor Cells of the Distal Lung: Impact on Fgfr2b-Driven Epithelial Repair. PLoS Pathogens, 12(6), e1005544.
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3

Cellular Stress Response Pathway Analysis

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Tunicamycin (TN), thapsigargin (TG), mouse anti-vimentin Cy 3 conjugated (#C5992), and mouse anti-pan cytokeratin FITC conjugated antibody (#C9080) were obtained from Sigma-Aldrich (St. Louis, MO). Anti-HGF affinity purified polyclonal antibody (#AF-294-SP) and recombinant HGF was purchased from R&D Systems (Abington U.K), and recombinant TGF-β1 was from MyBiosource Inc, (San Diego, USA).
Nita I., Hostettler K., Tamo L., Medová M., Bombaci G., Zhong J., Allam R., Zimmer Y., Roth M., Geiser T, & Gazdhar A. (2017). Hepatocyte growth factor secreted by bone marrow stem cell reduce ER stress and improves repair in alveolar epithelial II cells. Scientific Reports, 7, 41901.
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4

Analyzing MET and EGFR Signaling Pathways

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Primary antibodies used were anti-MET, anti-pMET, anti-EGFR, anti-pEGFR, anti-GAB1, anti-pGAB1, anti-AKT, anti-pAKT, anti-pERK1/2, (Cell Signaling Technology) and anti-β-actin (Sigma). The antibody arrays used were Proteome Profiler Human Phospho-Receptor Tyrosine Kinase (RTK) (ARY001B) and Phospho-Kinase Array (ARY003B) kits from R&D Systems. Recombinant HGF and pro-HGF were purchased from R&D Systems. The MET kinase inhibitor, JNJ38877605 was purchased from Selleck Chemicals (Selleckchem). HGF-specific and nonspecific (NSP) siRNAs were purchased from Dharmacon.
Owusu B.Y., Thomas S., Venukadasula P., Han Z., Janetka J.W., Galemmo RA J.r, & Klampfer L. (2017). Targeting the tumor-promoting microenvironment in MET-amplified NSCLC cells with a novel inhibitor of pro-HGF activation. Oncotarget, 8(38), 63014-63025.
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5

3D Spheroid Formation and Imaging

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The 3D protocol described in [18 ] was used. Briefly, 1 × 104 cells were plated on 12 mm circular cover glasses coated with Matrigel Matrix Growth Factor Reduced (MMGFR) (BD Biosciences) with serum free Mammary Epithelial Cell Growth Medium (MEGM™) media (Lonza, Walkersville, MD) containing the BulletKit™ growth supplement (BPE, hydrocortisone, GA-1000, Insulin). An overlay of MEGM growth media containing 2% MMGFR was added 1 hr after seeding. Spheroids were allowed to form overnight before addition of 100 ng recombinant HGF (R&D Biosystems) or pro-HGF conditioned media for 24 hrs after which cells were stained with 5 uM Cell Tracker Orange (Invitrogen) for 45 min and Hoechst 33342 (Life Technologies Carlsbad, CA) for 5 min prior to imaging. Confocal images were acquired on the Zeiss LSM 780 scope at the Microscopy Imaging and Cytometry Resources Core at Wayne State University School of Medicine.
Zoratti G.L., Tanabe L.M., Hyland T.E., Duhaime M.J., Colombo É., Leduc R., Marsault E., Johnson M.D., Lin C.Y., Boerner J., Lang J.E, & List K. (2016). Matriptase regulates c-Met mediated proliferation and invasion in inflammatory breast cancer. Oncotarget, 7(36), 58162-58173.
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6

HGF-Induced Growth Assay in NIH3T3 Cells

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NIH3T3 cells stably expressing wild-type or Asp153Tyr mutant MET were plated in complete medium. After 24 h, the medium was replaced with serum-free medium and cells were treated with the indicated concentration of recombinant HGF (R&D Systems). Cell growth was measured after 3 d with the CellTiter-Glo luminescence cell viability kit (Promega) as described previously98 (link). Student's t test (two-tailed) was used for statistical analyses to compare treatment groups with GraphPad Prism 5.00. A P value of <0.05 was considered statistically significant (*P < 0.05 and **P < 0.01).
Durinck S., Stawiski E.W., Pavía-Jiménez A., Modrusan Z., Kapur P., Jaiswal B.S., Zhang N., Toffessi-Tcheuyap V., Nguyen T.T., Pahuja K.B., Chen Y.J., Saleem S., Chaudhuri S., Heldens S., Jackson M., Peña-Llopis S., Guillory J., Toy K., Ha C., Harris C.J., Holloman E., Hill H.M., Stinson J., Rivers C.S., Janakiraman V., Wang W., Kinch L.N., Grishin N.V., Haverty P.M., Chow B., Gehring J.S., Reeder J., Pau G., Wu T.D., Margulis V., Lotan Y., Sagalowsky A., Pedrosa I., de Sauvage F.J., Brugarolas J, & Seshagiri S. (2014). Spectrum of diverse genomic alterations define non–clear cell renal carcinoma subtypes. Nature genetics, 47(1), 13-21.
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7

Recombinant HGF Dose-Dependent Proliferation

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Recombinant HGF (R&D Systems, Minneapolis, MN) was added to Pan02 (n = 5 repeat experiments) and TGP-47 cells (n = 4 repeat experiments) plated as described above, at the following doses: 0, 0.1, 1, 10, and 100 ng/ml. After 24 h of growth, the MTT proliferation assay was performed as above.
Ziegler K.M., Considine R.V., True E., Swartz-Basile D.A., Pitt H.A, & Zyromski N.J. (2016). Adipocytes enhance murine pancreatic cancer growth via a hepatocyte growth factor (HGF)-mediated mechanism. International journal of surgery (London, England), 28, 179-184.
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