Journal: Scientific reports

Article Title: Fibroblast Growth Factor Binding Protein 3 (FGFBP3) impacts carbohydrate and lipid metabolism.

doi: 10.1038/s41598-018-34238-5

Figure Lengend Snippet: Figure 5. BP3 binds to endocrine FGFs, enhances FGF19 and FGF21 signal transduction and promotes FGFR4/FGF19 complex formation. (A) Domains and computational structure prediction for BP1 and BP3. The FGF-binding domains in BP1 and BP3 in the C-terminal portion, the heparin-binding domain and the conserved cysteins of BP1 are shown. The white arrows in the 3D model indicate distinct folding of the FGF- binding domains. The PHYRE2 program (http://www.sbg.bio.ic.ac.uk/phyre/) was used68. (B) Binding of MBP-tagged BP1 or BP3 or MBP control to immobilized FGF19 measured by direct ELISA with an anti-MBP antibody. Mean ± SEM of one of three independent experiments done in duplicate (**P < 0.01; ***P < 0.0001). (C) SPR sensorgrams illustrating the binding of BP3 to immobilized FGF19. The concentrations of the BP3 analyte are indicated. RU, response units. (D) Binding of BP3 or MBP control to immobilized FGF15, or FGF23 measured by direct ELISA with an anti-MBP antibody. Mean ± SEM of one of three independent experiments done in duplicate. *P < 0.05 BP3 (black bars) vs. control (white bars). (E,F) Equilibrium binding of BP3 to immobilized FGF2 (E) or FGF19 (F) measured by direct ELISA with an anti-MBP antibody. Mean ± SEM of one of three independent experiments done in duplicate (**P < 0.01; ***P < 0.0001; Two-way ANOVA). (G) Competition of KLB and KLa proteins for binding of BP3 to immobilized FGF19 (left panel) or FGF21 (right panel). Mean ± SEM of one of three independent experiments run in duplicate (*P < 0.05; **P < 0.01 BP3 + KLB (red or blue dot) vs. BP3 + KLa (black diamond); Two-way ANOVA). (H) Schematic representation

Article Snippet: MaxiSorp microtiter plates were coated with 0.75 mg of human recombinant FGFR4 Fc Chimera (R&D Systems) and incubated overnight at 4 °C.

Techniques: Transduction, Binding Assay, Control, Direct ELISA

Journal: Investigative Ophthalmology & Visual Science

Article Title: Goblet Cell Differentiation Potential in Human Corneal Limbal Epithelial Progenitor Cells In Vitro

doi: 10.1167/iovs.61.12.27

Figure Lengend Snippet: FGFR, Fibroblast Growth Factor Receptor

Article Snippet: The following Abs were used: mouse anti-cytokeratin-3/2p monoclonal antibody (mAb; AE-5, 1:100; Santa Cruz Biotechnology, Inc, Dallas, TX, USA), mouse anti-human cytokeratin-4 mAb (6B10, 1:300; Abcam, Cambridge, UK), rabbit monoclonal [EP1599Y] anti-human cytokeratin 4 (1:200, Abcam), mouse anti-human cytokeratin-7 mAb (RCK105, 1:10000; Millipore, Billerica, MA, USA), mouse anti-human cytokeratin-13 mAb (1:20; American Research Products, Inc., Palos Verdes, CA, USA), mouse anti-human cytokeratin-12 mAb (N-16; 1:500; Santa Cruz Biotechnology), rabbit anti-cytokeratin 12/K12 mAb (1:200, EPR17882, Abcam), rabbit anti-human MUC5AC polyclonal Ab (H-160, 1:100, Santa Cruz Biotechnology), anti-FGFR1 mAb (1:200, 133105; R&D Systems, Minneapolis, MN, USA), anti-human FGFR2 (blocking) mAb (1:200, 98725; R&D Systems), anti-FGFR3 mAb (1:200, 135334; R&D Systems), anti-FGFR4 mAb (1:200, 137114; R&D Systems), and anti-FGFR1 blocking mAb (VBS1; Chemicon, Temecula, CA, USA).

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Goblet Cell Differentiation Potential in Human Corneal Limbal Epithelial Progenitor Cells In Vitro

doi: 10.1167/iovs.61.12.27

Figure Lengend Snippet: FGF receptor expression and effect of FGF receptor blockade in colonies derived from adherent single cells. ( A ) FGFR1 and FGFR2, but not FGFR3 and FGFR4, are detected in the adherent colonies by RT-PCR. ( B ) FGFR1 and FGFR2 expression is detected by Western blotting. ( C ) Effect of FGF receptor blockade was tested in colonies derived from adherent single cells. PAS-positive colony number in the anti-FGFR1 monoclonal antibody group significantly decreases as compared with that in the control Ig group. Treatment with an anti-FGFR2 blocking mAb does not affect the number of colony. ( D ) Colony with amorphous material and epithelium is observed with control IgG. ( E ) Anti-FGFR1 mAb suppresses the proliferation of both goblet-like cell and epithelium. Similar results were obtained with repeated three experiments. M, size markers; S, sample; P, positive control; N, negative control, N.S., not significant.

Article Snippet: The following Abs were used: mouse anti-cytokeratin-3/2p monoclonal antibody (mAb; AE-5, 1:100; Santa Cruz Biotechnology, Inc, Dallas, TX, USA), mouse anti-human cytokeratin-4 mAb (6B10, 1:300; Abcam, Cambridge, UK), rabbit monoclonal [EP1599Y] anti-human cytokeratin 4 (1:200, Abcam), mouse anti-human cytokeratin-7 mAb (RCK105, 1:10000; Millipore, Billerica, MA, USA), mouse anti-human cytokeratin-13 mAb (1:20; American Research Products, Inc., Palos Verdes, CA, USA), mouse anti-human cytokeratin-12 mAb (N-16; 1:500; Santa Cruz Biotechnology), rabbit anti-cytokeratin 12/K12 mAb (1:200, EPR17882, Abcam), rabbit anti-human MUC5AC polyclonal Ab (H-160, 1:100, Santa Cruz Biotechnology), anti-FGFR1 mAb (1:200, 133105; R&D Systems, Minneapolis, MN, USA), anti-human FGFR2 (blocking) mAb (1:200, 98725; R&D Systems), anti-FGFR3 mAb (1:200, 135334; R&D Systems), anti-FGFR4 mAb (1:200, 137114; R&D Systems), and anti-FGFR1 blocking mAb (VBS1; Chemicon, Temecula, CA, USA).

Techniques: Expressing, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Blocking Assay, Positive Control, Negative Control

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Schematic representation of the amperometric sensor for FGFR4 determination using magnetic immunocarriers and a sandwich format.

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques:

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Optimization of the experimental variables tested, according to the measured S/B ratio in the absence and in the presence of 5,000 pg mL -1 FGFR4 standard solutions, for the preparation of the amperometric immunosensor for FGFR4.

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Incubation

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Results are shown in the absence (white bars) or in the presence of 5,000 pg mL -1 FGFR4 (grey bars) together with the corresponding S/B ratio ( • ). 2(A), two sequential steps involving 30 min incubation of the CAb-MBs in a mixture solution containing FGFR4 and BDAb, and 30 min incubation in the Strep-HRP solution; 2(B) two steps involving 30 min incubation of the Cab-MBs with FGFR4 solution, followed by a 30 min incubation step in a mixture solution containing BDAb and Strep-HRP. Error bars estimated as triple of the standard deviation (n = 3).

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Incubation, Standard Deviation

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Amperometric signals measured in the absence (white bars) and in the presence of 2,500 pg mL -1 FGFR4 (grey bars) and the corresponding S/B ratio ( • ) in the absence (1) and in the presence of 10 ng mL -1 TNFα (2), 200 ng mL -1 human p53 (3), 5 mg mL -1 BSA (4), 5 mg mL -1 hemoglobin (5) and 1 mg mL -1 human IgG (6). Error bars estimated as triple of the standard deviation (n = 3).

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Standard Deviation

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Analysis of FGFR4 in cell lysates by Western Blot (10 μg) and amperometric traces recorded with the developed immunosensor (2.5 μg).

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Determination of endogenous FGFR4 concentration (in pg μg -1 ) in different cancer cell lysates (2.5 μg). Comparison of the results provided by the developed amperometric immunosensor with those obtained using a commercial ELISA spectrophotometric kit by performing three different determinations over the same cell lysate.

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Concentration Assay, Comparison, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Schematic representation of the amperometric sensor for FGFR4 determination using magnetic immunocarriers and a sandwich format.

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques:

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Optimization of the experimental variables tested, according to the measured S/B ratio in the absence and in the presence of 5,000 pg mL -1 FGFR4 standard solutions, for the preparation of the amperometric immunosensor for FGFR4.

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Incubation

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Results are shown in the absence (white bars) or in the presence of 5,000 pg mL -1 FGFR4 (grey bars) together with the corresponding S/B ratio ( • ). 2(A), two sequential steps involving 30 min incubation of the CAb-MBs in a mixture solution containing FGFR4 and BDAb, and 30 min incubation in the Strep-HRP solution; 2(B) two steps involving 30 min incubation of the Cab-MBs with FGFR4 solution, followed by a 30 min incubation step in a mixture solution containing BDAb and Strep-HRP. Error bars estimated as triple of the standard deviation (n = 3).

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Incubation, Standard Deviation

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Amperometric signals measured in the absence (white bars) and in the presence of 2,500 pg mL -1 FGFR4 (grey bars) and the corresponding S/B ratio ( • ) in the absence (1) and in the presence of 10 ng mL -1 TNFα (2), 200 ng mL -1 human p53 (3), 5 mg mL -1 BSA (4), 5 mg mL -1 hemoglobin (5) and 1 mg mL -1 human IgG (6). Error bars estimated as triple of the standard deviation (n = 3).

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Standard Deviation

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Analysis of FGFR4 in cell lysates by Western Blot (10 μg) and amperometric traces recorded with the developed immunosensor (2.5 μg).

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates

doi: 10.1371/journal.pone.0175056

Figure Lengend Snippet: Determination of endogenous FGFR4 concentration (in pg μg -1 ) in different cancer cell lysates (2.5 μg). Comparison of the results provided by the developed amperometric immunosensor with those obtained using a commercial ELISA spectrophotometric kit by performing three different determinations over the same cell lysate.

Article Snippet: Mouse anti-human FGFR4 antibody (CAb), biotinylated rat anti-human FGFR4 antibody (BDAb), and recombinant human FGFR4, as components of the Human Total FGFR4 DuoSet ® IC ELISA kit from (from R and D Systems, Inc., Minnneapolis, MN, USA, Catalog Number DYC685-2) were used.

Techniques: Concentration Assay, Comparison, Enzyme-linked Immunosorbent Assay