Recombinant bovine FGF2, human FGF9, human TGFβ1, mouse noggin/Fc chimera, human BMP4, human BMP6, and anti-BMP antibody (MAB3552) were from R&D Systems (Minneapolis, MN). The following antibodies were all purchased from Cell Signaling Technology (Danvers, MA): anti–phospho-p44/42 MAPK E10 mouse monoclonal (#9106), anti–total p44/42 MAPK (#9102), anti–phospho-p38 (#9211), anti–phospho-MEK1/2 (#9154), anti–phospho Raf-1 (#9427), anti–phospho-FRS2-α(Tyr-196) (#3864), and anti–phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465) (#9511). Other antibodies used were as follows: for CP49, rabbit anti-mouse CP49 polyclonal serum (#899 or #900; both generous gifts of Paul FitzGerald, University of California, Davis, CA); for phospho-tyrosine, 4G10 (a kind gift from Brian Druker, Oregon Health and Science University, Portland, OR); for luciferase, #G745A from Promega (Madison, WI); for GFP, JL-8 from Clontech (Mountain View, CA); for phospho-Smad3, ab51451 from Abcam (Cambridge, MA); for total Smad 1/5, ab75273 from Abcam; for total Raf-1, sc-7267 from Santa Cruz Biotechnology (Santa Cruz, CA); for total p38, sc-535 from Santa Cruz; and for total MEK, M17030 from Transduction Labs (Lexington, KT). UO126 (used at 15 μM), PD173074 (100 nM), and dorsomorphin (5 μM) were from Calbiochem (La Jolla, CA). All other reagents, including TPA, were from Sigma-Aldrich (St. Louis, MO).
Human fgf9
Manufactured by R&D Systems
Sourced in United States
Human FGF9 is a recombinant human fibroblast growth factor 9 protein. FGF9 is a member of the fibroblast growth factor family and is involved in various cellular processes such as cell growth, differentiation, and angiogenesis.
Automatically generated - may contain errors
Lab products found in correlation
Anti phospho mek1 2, by Cell Signaling Technology (1 mentions)
Ab51451, by Abcam (1 mentions)
Ab75273, by Abcam (1 mentions)
Human tgf β1, by R&D Systems (1 mentions)
Sc 535, by Santa Cruz Biotechnology (1 mentions)
Human bmp4, by R&D Systems (1 mentions)
Human bmp6, by R&D Systems (1 mentions)
Anti total p44 42 mapk, by Cell Signaling Technology (1 mentions)
Anti phospho frs2 α tyr196, by Cell Signaling Technology (1 mentions)
Sc 7267, by Santa Cruz Biotechnology (1 mentions)
Anti phospho p38, by Cell Signaling Technology (1 mentions)
Nitroblue tetrazolium, by Merck Group (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
Phospho p38, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Antibody against β actin, by Merck Group (1 mentions)
Sp600125, by Merck Group (1 mentions)
Sb203580, by Merck Group (1 mentions)
4 protocols using human fgf9
1
Investigating Intracellular Signaling Pathways
2
Signaling Pathway Analysis in Cell Culture
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Human FGF9 was purchased from R&D Systems (Minneapolis, MN, USA). Scintillation fluid was purchased from PerkinElmer (Boston, MA, USA). Fetal bovine serum and M199 powder were purchased from Gibco (Grand Island, NY, USA). Nitroblue tetrazolium, β-nicotinamide adenine dinucleotide, dihydroepiandrosterone, EDTA, ethylene glycol tetraacetic acid, sodium pyrophosphate, β-glycerophosphate and sodium orthovanadate were purchased from Sigma (Seelze, Germany). Waymouth's MB752/1, PD98059, SP600125, SB203580, and wortmannin were purchased from Sigma (St. Louis, MO, USA). 1,2,6,7-3H(N)-testosterone (70.00 Ci/mmol) and 1,2,6,7-3H(N)-progesterone (70.00 Ci/mmol) in 0.25 ml ethanol were purchased from DuPont-New England Nuclear (Boston, MA, USA). Antibodies against phospho-ERK1/2, phospho-p38, phospho-JNK, and phospho-Akt were purchased from Cell Signaling (Beverly, MA, USA). Antibody against β-actin was purchased from Sigma (St. Louis, MO, USA).
3
Nephron Progenitor Induction from iPSCs
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Three RCS-iPSCs and three control human iPS cell lines were induced toward nephron progenitors as previously described9 (link), with a minor modification. Briefly, human iPSCs were aggregated at 5 × 104 cells in V-bottomed 96-well low-cell-binding plates (Sumitomo Bakelite Co., Ltd.) using the medium containing 10 μM Y27632 and 0.5 ng/mL human bone morphogenic protein 4 (BMP4, R&D System). After 24 h (on day 1), the medium was changed to one containing 1 ng/mL human activin A (R&D System) and 20 ng/mL human fibroblast growth factor 2 (FGF2, R&D System). On day 3, the medium was changed to one containing 1 ng/mL human BMP4 and 10 μM CHIR99021 (Wako). On day 9, the medium was changed to one containing 10 ng/mL activin A, 3 ng/mL BMP4, 3 μM CHIR99021, 0.1 μM retinoic acid (Sigma), and 10 μM Y27632. On day 11, the medium was changed to one containing 1 μM CHIR99021, 5 ng/mL human FGF9 (R&D System), and 10 μM Y27632. On day 14, induced aggregates were cultured with NIH3T3 fibroblast expressing Wnt4 cells36 (link) supplied with DMEM containing 10% fetal bovine serum (FBS).
4
FGFR3IIIc Binding and Blocking Assay
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Various amounts of phage, Fab, or monoclonal antibody were serially diluted in 0.2% Tween 20/PBS containing 1% BSA, and added to hFGFR3IIIc-coated plates (50 µl at 1 µg/ml) and incubated at room temperature for 2 h. The bound antibodies were detected with an anti-human Fab antibody conjugated with HRP (Jackson ImmunoResearch, West Grove, PA; #109-035-097) in binding assays, or continuously probed with SULFO-TAG-labeled FGFs (Meso Scale Discovery, Rockville, MD; # R91AN-1) in blocking assays according to the supplier's instructions. 250 nM NaCl was present in ELISA binding and washing steps, and 10 µg/ml heparin was present in ligand-blocking assays. Human FGF1 (Genway Biotech Inc., San Diego, CA; #GWB-54AEB0), Human FGF9 (R&D Systems; #273-F9).
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