Anti fgfr3

Manufactured by Santa Cruz Biotechnology

Anti-FGFR3 is a laboratory reagent used in research applications. It is an antibody that specifically binds to the Fibroblast Growth Factor Receptor 3 (FGFR3) protein. This antibody can be utilized in various techniques, such as Western blotting, immunohistochemistry, and immunoprecipitation, to detect and study the FGFR3 protein.

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FGFR3 (15 citations)

6 protocols using anti fgfr3

Antibody Panel for Receptor Tyrosine Kinases

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Mouse monoclonal antibodies: anti-phospho-FAK(Tyr397) from BD Transduction (Lexington, KY); anti-EGFR and anti-phosphotyrosine clone 4G10, from Upstate Biotechnology (Lake Placid, NY); anti-β tubulin and anti-ERBB3 from Sigma-Aldrich (St. Louis MO); anti IGF-IRβ from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit polyclonal antibodies: anti-actin and anti-ERBB4 from Sigma; anti-phospho-FGFR (Tyr653/654), anti-phospho-HER2/ERBB2 (Tyr877), anti-phospho-HER4/ERBB4 (Tyr984) and anti-FAK from Cell Signaling (Beverly, MA); anti-PDGFR β and anti-PDGFRα from Upstate Biotechnology; anti-FGFR3 and anti-FGFR4 from Santa Cruz Biotechnology. Rabbit monoclonal antibodies: anti-phospho-PDGFRα (Tyr849)/PDGFRβ (Tyr857), anti-phospho-IGF-IRβ (Tyr1135/1136)/InsulinRβ (Tyr1150/1151), anti-phospho-HER3/ErbB3 (Tyr1289), anti-PDGFRβ and anti-HER2/ERBB2 from Cell Signaling.

Cassinelli G., Favini E., Dal Bo L., Tortoreto M., De Maglie M., Dagrada G., Pilotti S., Zunino F., Zaffaroni N, & Lanzi C. (2016). Antitumor efficacy of the heparan sulfate mimic roneparstat (SST0001) against sarcoma models involves multi-target inhibition of receptor tyrosine kinases. Oncotarget, 7(30), 47848-47863.

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Western Blot Analysis of FGF-2 Signaling

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The left cerebral hemisphere was separated and used. Equal amounts of protein samples (30μg) were loaded on SDS-PAGE gels, electrophoresed, and transferred onto a polyvinylidene difluoride membrane. The membranes were blocked with 5% non-fat blocking grade milk for 1 hour at room temperature followed by incubation overnight at 4 °C with following antibodies: anti-FGF-2 (1:100; sc-74412), anti-FGFR3 (1:100; sc-390423), anti-β-actin (1:5000; sc-47778) (Santa Cruz Biotechnology Inc., Dallas, TX), anti-FGFR1 (1:1000; 9740s), anti-PI3k (1:1000; 4249s), anti-p-Akt (1:1000; 9271s), anti-Akt (1:1000; 9272s) (Cell Signaling Technology Inc., Danvers, MA), anti-phosphorylated-FGFR (p-FGFR; 1:1000; PA5-64626, Thermo Fisher Scientific Inc., Waltham, MA), anti-Bax (1:1000; ab182734), and anti-Bcl-2 antibody(1:2500; ab59348, Abcam Inc., Cambridge, UK). On the following day, the membranes were incubated with the appropriate secondary antibody (1:5000; sc-516102, Santa Cruz Biotechnology Inc., Dallas, TX) and (1:5000; 12-348, MilliPore sigma Inc., Temecura, CA) at room temperature for 2 h. Immunoreactive bands were detected with a chemiluminescence reagent kit (ECL Prime; Amersham Biosciences Inc., Arlington Heights, IL) and quantified by densitometry with Image J software (NIH, Bethesda, MD).

Okada T., Enkhjargal B., Travis Z.D., Ocak U., Tang J., Suzuki H, & Zhang J.H. (2019). FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage. Molecular neurobiology, 56(12), 8203-8219.

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Inhibition of Crosstalk Signaling Pathways

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FGF-2 and BMP-2 were purchased from R&D Systems, chloroquine and PD173074 were from Sigma. MG132 was obtained from Calbiochem. Antibodies were from various sources, including Anti-Actin (AC-15) and Anti-Flag M2 from Sigma, anti-Myc (9E10), anti-HA (F-7), anti-GFP (FL), anti-FGFR3, anti-BMPR1b, and anti-PCNA from Santa Cruz, anti-pSmad1/5, anti-Smad1, and anti-Smad5 from Cell Signal Corporation, anti-Smurf1 antibody was from Invitrogen and anti-BMPR1a antibody from R&D Systems. Anti-pSmad1S214 was kindly provided by Eddy De Robertis [51 (link)].

Qi H., Jin M., Duan Y., Du X., Zhang Y., Ren F., Wang Y., Tian Q., Wang X., Wang Q., Zhu Y., Xie Y., Liu C., Cao X., Mishina Y., Chen D., Deng C.X., Chang Z, & Chen L. (2014). FGFR3 induces degradation of BMP type I receptor to regulate skeletal development. Biochimica et biophysica acta, 1843(7), 1237-1247.

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Immunofluorescence Staining of Neural Markers

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The following primary antibodies were used in this study: the monoclonal antibodies were: anti-βIII tubulin (1:500: mouse IgG, clone SDL3D10; Sigma-Aldrich), anti-FGFr3 (1:150: rabbit IgG; Santa Cruz Biotechnology), anti-GFAP (1:150: mouse IgG, clone GA5; Sigma-Aldrich), anti-nestin (1:500: mouse IgG; Millipore), anti-O4 (1:30; mouse IgM) (Sommer and Schachner, 1981 (link)), anti-vimentin (1:300: mouse IgG, clone LN-6; Sigma-Aldrich). The polyclonal antibodies were: anti-GFAP (1:300: rabbit IgG; Dako, Hamburg, Germany), anti-S100 (1:300: rabbit IgG; Dako). The specific secondary antibodies used in this study were CY2- (1:300) or CY3-coupled (1:500) anti-mouse and anti-rabbit antibodies (all from Dianova, Hamburg, Germany).

May M., Denecke B., Schroeder T., Götz M, & Faissner A. (2018). Cell tracking in vitro reveals that the extracellular matrix glycoprotein Tenascin-C modulates cell cycle length and differentiation in neural stem/progenitor cells of the developing mouse spinal cord. Biology Open, 7(7), bio027730.

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Western Blot Analysis of Cellular Signaling

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A total of 800,000 cells were seeded per well of a 6-well plate. Following overnight incubation, cells were treated with the indicated treatments and incubated overnight. Cells were washed and lysed using RIPA buffer (Cell Signaling Technology; #9806) freshly supplemented with phenylmethylsulfonyl fluoride (PMSF; Carl Roth; #329–98–6). Protein concentration was determined using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific; #23225) according to the manufacturer's instructions. Proteins were separated using SDS-PAGE, transferred to nitrocellulose membranes, then blotted with polyclonal anti-ERK1/2 (#9102), anti-pERK1/2 (clone 197G2; #4377), anti-AKT (clone 40D4; #2920), anti-pAKT (clone D9E; #4060), anti-FGFR3 (Santa Cruz Biotechnology; clone B-9; #sc-13121), anti-HER3 (clone D22C5; #12708), anti-pHER3 (clone 21D3; #4791), anti-β-actin (Sigma-Aldrich; clone AC-74; #A2228), polyclonal anti-Heregulin (#2573), polyclonal anti-EGFR (Santa Cruz Biotechnology; #sc-03), anti-pEGFR (clone 1H12, #2236), anti- HER2 (D8F12, #4290), anti-pHER2 (clone 21D3, #2247). All Western blot antibodies were purchased from Cell Signaling Technology unless otherwise specified.

Hosni S., Kilian V., Klümper N., Gabbia D., Sieckmann K., Corvino D., Winkler A., Saponaro M., Wörsdörfer K., Schmidt D., Hahn O., Zanotto I., Bertlich M., Toma M., Bald T., Eckstein M., Hölzel M., Geyer M., Ritter M., Wachten D., De Martin S, & Alajati A. (2024). Adipocyte Precursor-Derived NRG1 Promotes Resistance to FGFR Inhibition in Urothelial Carcinoma. Cancer Research, 84(5), 725-740.

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Immunofluorescence Analysis of FGFR-3 Expression

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Purified PCSCs at a density of 5×10⁵ cells/well were plated in six-well-plates, fixed with 4% paraformaldehyde, and permeabilized with 0.2% Triton X-100. Cells were then incubated with anti-FGFR-3 (1:1000 dilution, Santa Cruz) at 4 ºC overnight. Cells were then incubated with a FITC-labeled secondary antibody (Cellular Signaling Tech, Shanghai, China) for 1 h at room temperature. Cells were then observed and images recorded under a fluorescence microscope (Olympus, Tokyo, Japan).

Zhang F.Y., Zhen Y.F., Guo Z.X., Dai J., Zhu L.Q., Liang P.R., Su G.H., Zhang W.Y., Fang J.F., Yuan Q.W., Yao F., Liu Y., Qiao Y., Zhang Y., Guo W.L., Liu Y, & Wang X.D. (2021). miR-143 is implicated in growth plate injury by targeting IHH in precartilaginous stem cells. International Journal of Medical Sciences, 18(9), 1999-2007.

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