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biotinylated anti recombinant human fgf2 antibody  (R&D Systems)


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    R&D Systems biotinylated anti recombinant human fgf2 antibody
    Endothelial cells (BME-1) were cultured on Matrigel®-coated surfaces and incubated either without (A, C and E) or with the addition of <t>FGF2</t> and VEGF-165 (B, D and F) for 3 hrs. Panels A and B show phase contrast images, panels C and D show computer-generated projections of panels A and B used for quantification and panels E and F show the superimposed images. Panel G shows quantification of tube length (mean ± SEM of triplicate determinations in each case) as indicated. *** p < .001 relative to cells receiving no growth factors (left four bars) or to cells receiving growth factors without HIP/RPL29 (right three bars).
    Biotinylated Anti Recombinant Human Fgf2 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/biotinylated+anti+recombinant+human+fgf2+antibody/Human+FGF+basic%2FFGF2%2FbFGF+Biotinylated+Antibody/pmc04287213-154-21-26
    Average 90 stars, based on 7 article reviews
    biotinylated anti recombinant human fgf2 antibody - by Bioz Stars, 2026-10
    90/100 stars

    Images

    1) Product Images from "HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses"

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses

    Journal: Journal of cellular biochemistry

    doi: 10.1002/jcb.21899

    Endothelial cells (BME-1) were cultured on Matrigel®-coated surfaces and incubated either without (A, C and E) or with the addition of FGF2 and VEGF-165 (B, D and F) for 3 hrs. Panels A and B show phase contrast images, panels C and D show computer-generated projections of panels A and B used for quantification and panels E and F show the superimposed images. Panel G shows quantification of tube length (mean ± SEM of triplicate determinations in each case) as indicated. *** p < .001 relative to cells receiving no growth factors (left four bars) or to cells receiving growth factors without HIP/RPL29 (right three bars).
    Figure Legend Snippet: Endothelial cells (BME-1) were cultured on Matrigel®-coated surfaces and incubated either without (A, C and E) or with the addition of FGF2 and VEGF-165 (B, D and F) for 3 hrs. Panels A and B show phase contrast images, panels C and D show computer-generated projections of panels A and B used for quantification and panels E and F show the superimposed images. Panel G shows quantification of tube length (mean ± SEM of triplicate determinations in each case) as indicated. *** p < .001 relative to cells receiving no growth factors (left four bars) or to cells receiving growth factors without HIP/RPL29 (right three bars).

    Techniques Used: Cell Culture, Incubation, Generated

    Mouse aortic outgrowth assays were performed and quantified by computer-based morphometric analyses as described in Materials and Methods. Panels A–F show 8 day outgrowths from cultures grown in the presence of EBM with the following additions: A, none (control); B. 40 ng/ml FGF2; C, 40 ng/ml VEGF-165; D, 40 μg/ml HIP/RPL29; E, 40 ng/ml FGF2 plus 40 μg/ml HIP/RPL29; F, 40 ng/ml VEGF-165 plus 40 μg/ml HIP/RPL29. Panel G shows the quantitation of results of these types of assays and demonstrates near complete inhibition of outgrowth in the presence of HIP/RPL29 in all cases. * p < 0.001 vs. corresponding growth factor treatment in the absence of HIP/RPL29.
    Figure Legend Snippet: Mouse aortic outgrowth assays were performed and quantified by computer-based morphometric analyses as described in Materials and Methods. Panels A–F show 8 day outgrowths from cultures grown in the presence of EBM with the following additions: A, none (control); B. 40 ng/ml FGF2; C, 40 ng/ml VEGF-165; D, 40 μg/ml HIP/RPL29; E, 40 ng/ml FGF2 plus 40 μg/ml HIP/RPL29; F, 40 ng/ml VEGF-165 plus 40 μg/ml HIP/RPL29. Panel G shows the quantitation of results of these types of assays and demonstrates near complete inhibition of outgrowth in the presence of HIP/RPL29 in all cases. * p < 0.001 vs. corresponding growth factor treatment in the absence of HIP/RPL29.

    Techniques Used: Control, Quantitation Assay, Inhibition

    FGF2 (A) or VEGF-165 (B) were preincubated with perlecan domain I in a solid phase assay, unbound growth factor rinsed off and the substrates subsequently incubated with the indicated concentrations of HIP/RPL29 (HIP; filled circles) or lysozyme (LYS; open circles) for 2 hr. The surface was rinsed again and bound growth factor determined by ELISA as described in Materials and Methods. The points indicate the means ± SEM of triplicate determinations from a representative experiment.
    Figure Legend Snippet: FGF2 (A) or VEGF-165 (B) were preincubated with perlecan domain I in a solid phase assay, unbound growth factor rinsed off and the substrates subsequently incubated with the indicated concentrations of HIP/RPL29 (HIP; filled circles) or lysozyme (LYS; open circles) for 2 hr. The surface was rinsed again and bound growth factor determined by ELISA as described in Materials and Methods. The points indicate the means ± SEM of triplicate determinations from a representative experiment.

    Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay

    FGF2 was bound to a solid phase perlecan domain I substrate as described in Materials and Methods and Figure 5. In panel A, complexes subsequently were incubated with HPSE at the indicated concentrations for 24 hr, rinsed to remove unbound/released FGF2 and bound FGF2 measured by ELISA as described in Materials and Methods. The negative control was the perlecan domain I substrate not exposed to FGF2. In panel B, 5 μg/ml HPSE was incubated in the presence of the FGF2 bound to perlecan domain I for 24 hr at pH 5 or pH 7.2 as indicated and FGF2 release assayed by ELISA as described in Materials and Methods. FGF2 that remained bound at either pH 5.0 or 7.2 in the absence of HPSE served as a buffer only control. The bars represent the means ± SEM of triplicate determinations in each case.
    Figure Legend Snippet: FGF2 was bound to a solid phase perlecan domain I substrate as described in Materials and Methods and Figure 5. In panel A, complexes subsequently were incubated with HPSE at the indicated concentrations for 24 hr, rinsed to remove unbound/released FGF2 and bound FGF2 measured by ELISA as described in Materials and Methods. The negative control was the perlecan domain I substrate not exposed to FGF2. In panel B, 5 μg/ml HPSE was incubated in the presence of the FGF2 bound to perlecan domain I for 24 hr at pH 5 or pH 7.2 as indicated and FGF2 release assayed by ELISA as described in Materials and Methods. FGF2 that remained bound at either pH 5.0 or 7.2 in the absence of HPSE served as a buffer only control. The bars represent the means ± SEM of triplicate determinations in each case.

    Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay, Negative Control, Control

    FGF2 was bound to a solid phase perlecan domain I substrate as described in the legend to Figure 5. This complex subsequently was incubated with HPSE (5 μg/ml) alone or in the presence of 0.5, 5 or 40 μg/ml HIP/RPL29 as indicated on the figure. After 24 hr, the surfaces were rinsed to remove unbound/released FGF2 and bound FGF2 measured by ELISA as described in Materials and Methods. The negative control was the perlecan domain I substrate not exposed to FGF2. The bars represent the means ± SEM of triplicate determinations from a representative experiment in each case.
    Figure Legend Snippet: FGF2 was bound to a solid phase perlecan domain I substrate as described in the legend to Figure 5. This complex subsequently was incubated with HPSE (5 μg/ml) alone or in the presence of 0.5, 5 or 40 μg/ml HIP/RPL29 as indicated on the figure. After 24 hr, the surfaces were rinsed to remove unbound/released FGF2 and bound FGF2 measured by ELISA as described in Materials and Methods. The negative control was the perlecan domain I substrate not exposed to FGF2. The bars represent the means ± SEM of triplicate determinations from a representative experiment in each case.

    Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay, Negative Control

    Related Articles

    Cell Culture:

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses
    Article Snippet: Following washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature.washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature. ... After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.. After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.

    Incubation:

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses
    Article Snippet: Following washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature.washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature. ... After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.. After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.

    Generated:

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses
    Article Snippet: Following washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature.washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature. ... After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.. After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.

    Control:

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses
    Article Snippet: Following washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature.washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature. ... After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.. After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.

    Quantitation Assay:

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses
    Article Snippet: Following washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature.washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature. ... After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.. After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.

    Inhibition:

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses
    Article Snippet: Following washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature.washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature. ... After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.. After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.

    Enzyme-linked Immunosorbent Assay:

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses
    Article Snippet: Following washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature.washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature. ... After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.. After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.

    Negative Control:

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses
    Article Snippet: Following washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature.washing three times with 0.05% (v/v) Tween 20 in PBS, 100 μl of HIP/RPL29 in blocking buffer was added at increasing concentrations (0, 0.05, 0.5, 5, 10, 20, 40 and 80g/ml) to each well of a 96-well microplate and incubated for 2 hours at room temperature. ... After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.. After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.After incubation with 0.1 μg/ml of horse radish peroxidase-conjugated NeutrAvidin (Pierce, Rockford, IL) in SuperBlock Blocking Buffer (Pierce, Rockford, IL) for 30 min, the wells were again rinsed several times as above and then reacted with 200 μl of TMB solution followed by washing with PBS.



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    biotinylated anti recombinant human fgf2 antibody  (R&D Systems)
    90
    R&D Systems biotinylated anti recombinant human fgf2 antibody
    Endothelial cells (BME-1) were cultured on Matrigel®-coated surfaces and incubated either without (A, C and E) or with the addition of <t>FGF2</t> and VEGF-165 (B, D and F) for 3 hrs. Panels A and B show phase contrast images, panels C and D show computer-generated projections of panels A and B used for quantification and panels E and F show the superimposed images. Panel G shows quantification of tube length (mean ± SEM of triplicate determinations in each case) as indicated. *** p < .001 relative to cells receiving no growth factors (left four bars) or to cells receiving growth factors without HIP/RPL29 (right three bars).
    Biotinylated Anti Recombinant Human Fgf2 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/biotinylated+anti+recombinant+human+fgf2+antibody/Human+FGF+basic%2FFGF2%2FbFGF+Biotinylated+Antibody/pmc04287213-154-21-26
    Average 90 stars, based on 1 article reviews
    biotinylated anti recombinant human fgf2 antibody - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Endothelial cells (BME-1) were cultured on Matrigel®-coated surfaces and incubated either without (A, C and E) or with the addition of FGF2 and VEGF-165 (B, D and F) for 3 hrs. Panels A and B show phase contrast images, panels C and D show computer-generated projections of panels A and B used for quantification and panels E and F show the superimposed images. Panel G shows quantification of tube length (mean ± SEM of triplicate determinations in each case) as indicated. *** p < .001 relative to cells receiving no growth factors (left four bars) or to cells receiving growth factors without HIP/RPL29 (right three bars).

    Journal: Journal of cellular biochemistry

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses

    doi: 10.1002/jcb.21899

    Figure Lengend Snippet: Endothelial cells (BME-1) were cultured on Matrigel®-coated surfaces and incubated either without (A, C and E) or with the addition of FGF2 and VEGF-165 (B, D and F) for 3 hrs. Panels A and B show phase contrast images, panels C and D show computer-generated projections of panels A and B used for quantification and panels E and F show the superimposed images. Panel G shows quantification of tube length (mean ± SEM of triplicate determinations in each case) as indicated. *** p < .001 relative to cells receiving no growth factors (left four bars) or to cells receiving growth factors without HIP/RPL29 (right three bars).

    Article Snippet: After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.

    Techniques: Cell Culture, Incubation, Generated

    Mouse aortic outgrowth assays were performed and quantified by computer-based morphometric analyses as described in Materials and Methods. Panels A–F show 8 day outgrowths from cultures grown in the presence of EBM with the following additions: A, none (control); B. 40 ng/ml FGF2; C, 40 ng/ml VEGF-165; D, 40 μg/ml HIP/RPL29; E, 40 ng/ml FGF2 plus 40 μg/ml HIP/RPL29; F, 40 ng/ml VEGF-165 plus 40 μg/ml HIP/RPL29. Panel G shows the quantitation of results of these types of assays and demonstrates near complete inhibition of outgrowth in the presence of HIP/RPL29 in all cases. * p < 0.001 vs. corresponding growth factor treatment in the absence of HIP/RPL29.

    Journal: Journal of cellular biochemistry

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses

    doi: 10.1002/jcb.21899

    Figure Lengend Snippet: Mouse aortic outgrowth assays were performed and quantified by computer-based morphometric analyses as described in Materials and Methods. Panels A–F show 8 day outgrowths from cultures grown in the presence of EBM with the following additions: A, none (control); B. 40 ng/ml FGF2; C, 40 ng/ml VEGF-165; D, 40 μg/ml HIP/RPL29; E, 40 ng/ml FGF2 plus 40 μg/ml HIP/RPL29; F, 40 ng/ml VEGF-165 plus 40 μg/ml HIP/RPL29. Panel G shows the quantitation of results of these types of assays and demonstrates near complete inhibition of outgrowth in the presence of HIP/RPL29 in all cases. * p < 0.001 vs. corresponding growth factor treatment in the absence of HIP/RPL29.

    Article Snippet: After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.

    Techniques: Control, Quantitation Assay, Inhibition

    FGF2 (A) or VEGF-165 (B) were preincubated with perlecan domain I in a solid phase assay, unbound growth factor rinsed off and the substrates subsequently incubated with the indicated concentrations of HIP/RPL29 (HIP; filled circles) or lysozyme (LYS; open circles) for 2 hr. The surface was rinsed again and bound growth factor determined by ELISA as described in Materials and Methods. The points indicate the means ± SEM of triplicate determinations from a representative experiment.

    Journal: Journal of cellular biochemistry

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses

    doi: 10.1002/jcb.21899

    Figure Lengend Snippet: FGF2 (A) or VEGF-165 (B) were preincubated with perlecan domain I in a solid phase assay, unbound growth factor rinsed off and the substrates subsequently incubated with the indicated concentrations of HIP/RPL29 (HIP; filled circles) or lysozyme (LYS; open circles) for 2 hr. The surface was rinsed again and bound growth factor determined by ELISA as described in Materials and Methods. The points indicate the means ± SEM of triplicate determinations from a representative experiment.

    Article Snippet: After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay

    FGF2 was bound to a solid phase perlecan domain I substrate as described in Materials and Methods and Figure 5. In panel A, complexes subsequently were incubated with HPSE at the indicated concentrations for 24 hr, rinsed to remove unbound/released FGF2 and bound FGF2 measured by ELISA as described in Materials and Methods. The negative control was the perlecan domain I substrate not exposed to FGF2. In panel B, 5 μg/ml HPSE was incubated in the presence of the FGF2 bound to perlecan domain I for 24 hr at pH 5 or pH 7.2 as indicated and FGF2 release assayed by ELISA as described in Materials and Methods. FGF2 that remained bound at either pH 5.0 or 7.2 in the absence of HPSE served as a buffer only control. The bars represent the means ± SEM of triplicate determinations in each case.

    Journal: Journal of cellular biochemistry

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses

    doi: 10.1002/jcb.21899

    Figure Lengend Snippet: FGF2 was bound to a solid phase perlecan domain I substrate as described in Materials and Methods and Figure 5. In panel A, complexes subsequently were incubated with HPSE at the indicated concentrations for 24 hr, rinsed to remove unbound/released FGF2 and bound FGF2 measured by ELISA as described in Materials and Methods. The negative control was the perlecan domain I substrate not exposed to FGF2. In panel B, 5 μg/ml HPSE was incubated in the presence of the FGF2 bound to perlecan domain I for 24 hr at pH 5 or pH 7.2 as indicated and FGF2 release assayed by ELISA as described in Materials and Methods. FGF2 that remained bound at either pH 5.0 or 7.2 in the absence of HPSE served as a buffer only control. The bars represent the means ± SEM of triplicate determinations in each case.

    Article Snippet: After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Negative Control, Control

    FGF2 was bound to a solid phase perlecan domain I substrate as described in the legend to Figure 5. This complex subsequently was incubated with HPSE (5 μg/ml) alone or in the presence of 0.5, 5 or 40 μg/ml HIP/RPL29 as indicated on the figure. After 24 hr, the surfaces were rinsed to remove unbound/released FGF2 and bound FGF2 measured by ELISA as described in Materials and Methods. The negative control was the perlecan domain I substrate not exposed to FGF2. The bars represent the means ± SEM of triplicate determinations from a representative experiment in each case.

    Journal: Journal of cellular biochemistry

    Article Title: HIP/RPL29 Antagonizes VEGF and FGF2 Stimulated Angiogenesis by Interfering with HS-dependent Responses

    doi: 10.1002/jcb.21899

    Figure Lengend Snippet: FGF2 was bound to a solid phase perlecan domain I substrate as described in the legend to Figure 5. This complex subsequently was incubated with HPSE (5 μg/ml) alone or in the presence of 0.5, 5 or 40 μg/ml HIP/RPL29 as indicated on the figure. After 24 hr, the surfaces were rinsed to remove unbound/released FGF2 and bound FGF2 measured by ELISA as described in Materials and Methods. The negative control was the perlecan domain I substrate not exposed to FGF2. The bars represent the means ± SEM of triplicate determinations from a representative experiment in each case.

    Article Snippet: After washing three times with 0.05% (v/v) Tween 20 in PBS, the bound FGF2 and VEGF-165 were identified with 2 μg/ml biotinylated anti-recombinant human FGF2 antibody (R&D Systems, BAM-233) and 3 μg/ml biotinylated anti-recombinant human VEGF antibody (R&D Systems, BAF-293), respectively.

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Negative Control