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R&D Systems anti hb egf neutralizing antibody

TNIIIA2 secreted SASP factors, including <t>HB-EGF,</t> induce the malignant transformation of HaCaT cells. In (A and B), HaCaT cells cocultured with TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF <t>neutralizing</t> antibody or isotype control. After incubation for 72 h, the HaCaT area was measured by rhodanile blue staining. Scale bar, 200 μm. (C) HaCaT cells treated with CM from TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control and then subjected to a WST-8 (D) or colony formation (E) assay. (F) WI-38 cells were treated with either TNIIIA2 (50 μg/mL) or NaB (4 mM) to induce cellular senescence, as described in the Materials and Methods. HB-EGF and GAPDH mRNA levels were then measured by semi-quantitative PCR. In (G and H), HaCaT cells were treated with recombinant human HB-EGF, and the cells were subjected to a WST-8 assay on day 2 (G) or colony formation assay on day 10 (H). Data are presented as means ± SD, *P < 0.05, **P < 0.01.

Anti Hb Egf Neutralizing Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 98 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 98 article reviews

anti hb egf neutralizing antibody - by Bioz Stars, 2026-07

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1) Product Images from "Induction of cellular senescence in fibroblasts through β1-integrin activation by tenascin-C-derived peptide and its protumor effect"

Article Title: Induction of cellular senescence in fibroblasts through β1-integrin activation by tenascin-C-derived peptide and its protumor effect

Journal: American Journal of Cancer Research

doi:

TNIIIA2 secreted SASP factors, including HB-EGF, induce the malignant transformation of HaCaT cells. In (A and B), HaCaT cells cocultured with TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control. After incubation for 72 h, the HaCaT area was measured by rhodanile blue staining. Scale bar, 200 μm. (C) HaCaT cells treated with CM from TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control and then subjected to a WST-8 (D) or colony formation (E) assay. (F) WI-38 cells were treated with either TNIIIA2 (50 μg/mL) or NaB (4 mM) to induce cellular senescence, as described in the Materials and Methods. HB-EGF and GAPDH mRNA levels were then measured by semi-quantitative PCR. In (G and H), HaCaT cells were treated with recombinant human HB-EGF, and the cells were subjected to a WST-8 assay on day 2 (G) or colony formation assay on day 10 (H). Data are presented as means ± SD, *P < 0.05, **P < 0.01.

Figure Legend Snippet: TNIIIA2 secreted SASP factors, including HB-EGF, induce the malignant transformation of HaCaT cells. In (A and B), HaCaT cells cocultured with TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control. After incubation for 72 h, the HaCaT area was measured by rhodanile blue staining. Scale bar, 200 μm. (C) HaCaT cells treated with CM from TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control and then subjected to a WST-8 (D) or colony formation (E) assay. (F) WI-38 cells were treated with either TNIIIA2 (50 μg/mL) or NaB (4 mM) to induce cellular senescence, as described in the Materials and Methods. HB-EGF and GAPDH mRNA levels were then measured by semi-quantitative PCR. In (G and H), HaCaT cells were treated with recombinant human HB-EGF, and the cells were subjected to a WST-8 assay on day 2 (G) or colony formation assay on day 10 (H). Data are presented as means ± SD, *P < 0.05, **P < 0.01.

Techniques Used: Transformation Assay, Incubation, Control, Staining, Real-time Polymerase Chain Reaction, Recombinant, Colony Assay

A model of malignant transformation of preneoplastic cells via factors secreted from TNIIIA2-induced senescent fibroblasts. Activation of β1-integrin by TNIIIA2 induces fibroblast senescence. Factors secreted by TNIIIA2-induced senescent fibroblasts, including HB-EGF, are involved in the malignant transformation of preneoplastic cells. Inactivation of β1-integrin by peptide FNIII14 inhibits TNIIIA2-induced cellular senescence. Created using BioRender.com.

Figure Legend Snippet: A model of malignant transformation of preneoplastic cells via factors secreted from TNIIIA2-induced senescent fibroblasts. Activation of β1-integrin by TNIIIA2 induces fibroblast senescence. Factors secreted by TNIIIA2-induced senescent fibroblasts, including HB-EGF, are involved in the malignant transformation of preneoplastic cells. Inactivation of β1-integrin by peptide FNIII14 inhibits TNIIIA2-induced cellular senescence. Created using BioRender.com.

Techniques Used: Transformation Assay, Activation Assay

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Journal: American Journal of Cancer Research

Article Title: Induction of cellular senescence in fibroblasts through β1-integrin activation by tenascin-C-derived peptide and its protumor effect

doi:

Figure Lengend Snippet: TNIIIA2 secreted SASP factors, including HB-EGF, induce the malignant transformation of HaCaT cells. In (A and B), HaCaT cells cocultured with TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control. After incubation for 72 h, the HaCaT area was measured by rhodanile blue staining. Scale bar, 200 μm. (C) HaCaT cells treated with CM from TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control and then subjected to a WST-8 (D) or colony formation (E) assay. (F) WI-38 cells were treated with either TNIIIA2 (50 μg/mL) or NaB (4 mM) to induce cellular senescence, as described in the Materials and Methods. HB-EGF and GAPDH mRNA levels were then measured by semi-quantitative PCR. In (G and H), HaCaT cells were treated with recombinant human HB-EGF, and the cells were subjected to a WST-8 assay on day 2 (G) or colony formation assay on day 10 (H). Data are presented as means ± SD, *P < 0.05, **P < 0.01.

Article Snippet: Recombinant human heparin-binding epidermal growth factor-like growth factor (HB-EGF) and an anti-HB-EGF neutralizing antibody (AF-259-NA) were obtained from R&D Systems (Minneapolis, MN).

Techniques: Transformation Assay, Incubation, Control, Staining, Real-time Polymerase Chain Reaction, Recombinant, Colony Assay

Journal: American Journal of Cancer Research

Article Title: Induction of cellular senescence in fibroblasts through β1-integrin activation by tenascin-C-derived peptide and its protumor effect

doi:

Figure Lengend Snippet: A model of malignant transformation of preneoplastic cells via factors secreted from TNIIIA2-induced senescent fibroblasts. Activation of β1-integrin by TNIIIA2 induces fibroblast senescence. Factors secreted by TNIIIA2-induced senescent fibroblasts, including HB-EGF, are involved in the malignant transformation of preneoplastic cells. Inactivation of β1-integrin by peptide FNIII14 inhibits TNIIIA2-induced cellular senescence. Created using BioRender.com.

Techniques: Transformation Assay, Activation Assay

The alterations in the gene expression in cells cultured 2, 6, and 24 h under different conditions were determined with real-time RT-PCR analysis. The following conditions were tested: hyperosmolarity induced by addition of NaCl (100 mM; A,B ) or sucrose (100 mM; C ) to the culture medium, hypoosmolarity (60% osmolarity; D ), chemical hypoxia induced by addition of CoCl 2 (150 μM; E ), and oxidative stress induced by addition of H 2 O 2 (20 μM; F ). ( B). Dose-dependent effect of high extracellular NaCl on the cellular levels of bFGF and HB-EGF mRNAs. The cells were cultured in media which were made up hyperosmotic by addition of 10 to 100 mM NaCl. Means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05.

Journal: PLoS ONE

Article Title: Osmotic Induction of Angiogenic Growth Factor Expression in Human Retinal Pigment Epithelial Cells

doi: 10.1371/journal.pone.0147312

Figure Lengend Snippet: The alterations in the gene expression in cells cultured 2, 6, and 24 h under different conditions were determined with real-time RT-PCR analysis. The following conditions were tested: hyperosmolarity induced by addition of NaCl (100 mM; A,B ) or sucrose (100 mM; C ) to the culture medium, hypoosmolarity (60% osmolarity; D ), chemical hypoxia induced by addition of CoCl 2 (150 μM; E ), and oxidative stress induced by addition of H 2 O 2 (20 μM; F ). ( B). Dose-dependent effect of high extracellular NaCl on the cellular levels of bFGF and HB-EGF mRNAs. The cells were cultured in media which were made up hyperosmotic by addition of 10 to 100 mM NaCl. Means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05.

Article Snippet: The following antibodies were used: a neutralizing goat anti-bFGF (R&D Systems), a neutralizing goat anti-HB-EGF (R&D Systems), a neutralizing rabbit anti-transforming growth factor (TGF)-β (pan specific; R&D Systems), a rabbit anti-human NFAT5 (1:200; Santa Cruz Biotechnology, Dallas TX), a rabbit anti-human β-actin (1:1000; Cell Signaling, Frankfurt/M., Germany), and anti-rabbit IgG conjugated with alkaline phosphatase (1:2000; Cell Signaling).

Techniques: Expressing, Cell Culture, Quantitative RT-PCR

A,B. Cellular levels of bFGF ( A ) and HB-EGF mRNAs ( B ) were determined with real-time RT-PCR analysis in cells cultured 6 h in iso- (control) and hyperosmotic (+ 100 mM NaCl) media, respectively. ( C ). The mRNA level was determined in cells cultured 24 h under control conditions and in the presence of CoCl 2 (150 μM), respectively. The following pharmacological inhibitors were tested: the inhibitor of p38α/β MAPK activation, SB203580 (10 μM), the inhibitor of ERK1/2 activation, PD98059 (20 μM), the JNK inhibitor SP600125 (10 μM), and the inhibitor of PI3K-related kinases, LY294002 (5 μM). The vehicle control was made with dimethylsulfoxide (DMSO; 1:1000). Means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05.

Journal: PLoS ONE

Article Title: Osmotic Induction of Angiogenic Growth Factor Expression in Human Retinal Pigment Epithelial Cells

doi: 10.1371/journal.pone.0147312

Figure Lengend Snippet: A,B. Cellular levels of bFGF ( A ) and HB-EGF mRNAs ( B ) were determined with real-time RT-PCR analysis in cells cultured 6 h in iso- (control) and hyperosmotic (+ 100 mM NaCl) media, respectively. ( C ). The mRNA level was determined in cells cultured 24 h under control conditions and in the presence of CoCl 2 (150 μM), respectively. The following pharmacological inhibitors were tested: the inhibitor of p38α/β MAPK activation, SB203580 (10 μM), the inhibitor of ERK1/2 activation, PD98059 (20 μM), the JNK inhibitor SP600125 (10 μM), and the inhibitor of PI3K-related kinases, LY294002 (5 μM). The vehicle control was made with dimethylsulfoxide (DMSO; 1:1000). Means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05.

Techniques: Quantitative RT-PCR, Cell Culture, Activation Assay

Cellular levels of bFGF ( A ), HB-EGF ( B ), and VEGF mRNAs ( D ) were measured with real-time RT-PCR analysis in cells cultured 6 h in iso- (control) and hyperosmotic (+ 100 mM NaCl) media. The hypoxic expression of the HB-EGF gene ( C ) was determined in cells cultured 24 h in the absence (control) and presence of CoCl 2 (150 μM). The following pharmacological inhibitors were tested: the inhibitor of the EGF receptor tyrosine kinase, AG1478 (600 nM), the inhibitor of TGF-β1 superfamily activin receptor-like kinase receptors, SB431542 (10 μM), the FGF receptor kinase inhibitor, PD173074 (500 nM), and the broad-spectrum metalloproteinase inhibitor 1,10-phenanthroline (1,10-Phen; 10 μM). Means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05.

Journal: PLoS ONE

Article Title: Osmotic Induction of Angiogenic Growth Factor Expression in Human Retinal Pigment Epithelial Cells

doi: 10.1371/journal.pone.0147312

Figure Lengend Snippet: Cellular levels of bFGF ( A ), HB-EGF ( B ), and VEGF mRNAs ( D ) were measured with real-time RT-PCR analysis in cells cultured 6 h in iso- (control) and hyperosmotic (+ 100 mM NaCl) media. The hypoxic expression of the HB-EGF gene ( C ) was determined in cells cultured 24 h in the absence (control) and presence of CoCl 2 (150 μM). The following pharmacological inhibitors were tested: the inhibitor of the EGF receptor tyrosine kinase, AG1478 (600 nM), the inhibitor of TGF-β1 superfamily activin receptor-like kinase receptors, SB431542 (10 μM), the FGF receptor kinase inhibitor, PD173074 (500 nM), and the broad-spectrum metalloproteinase inhibitor 1,10-phenanthroline (1,10-Phen; 10 μM). Means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05.

Techniques: Quantitative RT-PCR, Cell Culture, Expressing

mRNA levels in cells cultured in hyperosmotic (+ 100 mM NaCl) medium were measured with real-time RT-PCR analysis ( A,D,E ). The cellular protein levels ( B,C ) were determined by Western blot analysis. After transfection of the cells with NFAT5 siRNA (siNFAT5) or nontargeted siRNA (siNon; 50 nM each) for 24 h, the cells were cultured 16 h in serum-free isoosmotic control medium. Thereafter, serum-free hyperosmotic medium was added for 6 h. ( A ). Transfection with siNFAT5 resulted in a reduction of the NFAT5 mRNA level under hyperosmotic conditions. ( B ). Effect of siRNA transfection on the cellular level of NFAT5 protein under isoosmotic control conditions. ( C ). Effect of siRNA transfection on the cellular level of NFAT5 protein under hyperosmotic conditions. ( D ). Transfection with siNFAT5 reduced the level of bFGF mRNA in cells cultured in hyperosmotic medium compared to nontransfected cells. Nontargeted siRNA was without effect. ( E ). Effects of siNFAT5 and nontargeted siRNA on the level of HB-EGF mRNA in cells cultured in hyperosmotic medium. Bars are means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05. Significant difference vs . nontargeted siRNA: ° P <0.05. In B and C , 35 ( B ) and 60 μg ( C ) of total protein were used for separation. β-Actin was used as a control for equal protein loading. Similar results were obtained in 3 independent experiments using cells from different donors.

Journal: PLoS ONE

Article Title: Osmotic Induction of Angiogenic Growth Factor Expression in Human Retinal Pigment Epithelial Cells

doi: 10.1371/journal.pone.0147312

Figure Lengend Snippet: mRNA levels in cells cultured in hyperosmotic (+ 100 mM NaCl) medium were measured with real-time RT-PCR analysis ( A,D,E ). The cellular protein levels ( B,C ) were determined by Western blot analysis. After transfection of the cells with NFAT5 siRNA (siNFAT5) or nontargeted siRNA (siNon; 50 nM each) for 24 h, the cells were cultured 16 h in serum-free isoosmotic control medium. Thereafter, serum-free hyperosmotic medium was added for 6 h. ( A ). Transfection with siNFAT5 resulted in a reduction of the NFAT5 mRNA level under hyperosmotic conditions. ( B ). Effect of siRNA transfection on the cellular level of NFAT5 protein under isoosmotic control conditions. ( C ). Effect of siRNA transfection on the cellular level of NFAT5 protein under hyperosmotic conditions. ( D ). Transfection with siNFAT5 reduced the level of bFGF mRNA in cells cultured in hyperosmotic medium compared to nontransfected cells. Nontargeted siRNA was without effect. ( E ). Effects of siNFAT5 and nontargeted siRNA on the level of HB-EGF mRNA in cells cultured in hyperosmotic medium. Bars are means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05. Significant difference vs . nontargeted siRNA: ° P <0.05. In B and C , 35 ( B ) and 60 μg ( C ) of total protein were used for separation. β-Actin was used as a control for equal protein loading. Similar results were obtained in 3 independent experiments using cells from different donors.

Techniques: Cell Culture, Quantitative RT-PCR, Western Blot, Transfection

The level of bFGF protein was determined with ELISA in the media of cells cultured 6 h under iso- (control) and hyperosmotic (+ 100 mM NaCl) conditions. ( A ). The following pharmacological inhibitors were tested: the inhibitor of TGF-β1 superfamily activin receptor-like kinase receptors, SB431542 (10 μM), the FGF receptor kinase inhibitor, PD173074 (500 nM), the inhibitor of the EGF receptor tyrosine kinase, AG1478 (600 nM), neutralizing antibodies against TGF-β (anti-TGF-β) and HB-EGF (anti-HB-EGF), respectively (each at 20 μg/ml), as well as the broad-spectrum metalloproteinase inhibitor 1,10-phenanthroline (1,10-Phen; 10 μM). As control, a neutralizing antibody against bFGF (anti-bFGF; 20 μg/ml) was used. The vehicle control was made with dimethylsulfoxide (DMSO; 1:1000). ( B ). The hyperosmotic secretion of bFGF from cells transfected with NFTA5 siRNA (siNFAT5; 10 nM) was smaller than secretion of bFGF from cells transfected with nontargeted siRNA (siNon; 10 nM). Means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05. Significant difference vs . nontargeted siRNA: ° P <0.05.

Journal: PLoS ONE

Article Title: Osmotic Induction of Angiogenic Growth Factor Expression in Human Retinal Pigment Epithelial Cells

doi: 10.1371/journal.pone.0147312

Figure Lengend Snippet: The level of bFGF protein was determined with ELISA in the media of cells cultured 6 h under iso- (control) and hyperosmotic (+ 100 mM NaCl) conditions. ( A ). The following pharmacological inhibitors were tested: the inhibitor of TGF-β1 superfamily activin receptor-like kinase receptors, SB431542 (10 μM), the FGF receptor kinase inhibitor, PD173074 (500 nM), the inhibitor of the EGF receptor tyrosine kinase, AG1478 (600 nM), neutralizing antibodies against TGF-β (anti-TGF-β) and HB-EGF (anti-HB-EGF), respectively (each at 20 μg/ml), as well as the broad-spectrum metalloproteinase inhibitor 1,10-phenanthroline (1,10-Phen; 10 μM). As control, a neutralizing antibody against bFGF (anti-bFGF; 20 μg/ml) was used. The vehicle control was made with dimethylsulfoxide (DMSO; 1:1000). ( B ). The hyperosmotic secretion of bFGF from cells transfected with NFTA5 siRNA (siNFAT5; 10 nM) was smaller than secretion of bFGF from cells transfected with nontargeted siRNA (siNon; 10 nM). Means ± SEM of 3–7 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05. Significant difference vs . nontargeted siRNA: ° P <0.05.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Transfection

bFGF ( A ), HB-EGF ( B,D ), and NFAT5 ( C ) mRNA levels in cells cultured 6 ( A-C ) and 24 h ( D ) under hyperosmotic ( A-C ) and hypoxic conditions ( D ) were determined with real-time RT-PCR analysis. Hyperosmolarity was induced by addition of NaCl (100 mM) to the culture medium. Chemical hypoxia was induced by addition of CoCl 2 (150 μM). The following polyphenols were tested: myricetin (50 μM), cyanidin (100 μM), luteolin (50 μM), quercetin (100 μM), apigenin (50 μM), and EGCG (50 μM). Vehicle controls were made with acetone (0.1%), dimethylsulfoxide (DMSO; 0.1%), and methanol (0.2%). Means ± SEM of 3–6 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05.

Journal: PLoS ONE

Article Title: Osmotic Induction of Angiogenic Growth Factor Expression in Human Retinal Pigment Epithelial Cells

doi: 10.1371/journal.pone.0147312

Figure Lengend Snippet: bFGF ( A ), HB-EGF ( B,D ), and NFAT5 ( C ) mRNA levels in cells cultured 6 ( A-C ) and 24 h ( D ) under hyperosmotic ( A-C ) and hypoxic conditions ( D ) were determined with real-time RT-PCR analysis. Hyperosmolarity was induced by addition of NaCl (100 mM) to the culture medium. Chemical hypoxia was induced by addition of CoCl 2 (150 μM). The following polyphenols were tested: myricetin (50 μM), cyanidin (100 μM), luteolin (50 μM), quercetin (100 μM), apigenin (50 μM), and EGCG (50 μM). Vehicle controls were made with acetone (0.1%), dimethylsulfoxide (DMSO; 0.1%), and methanol (0.2%). Means ± SEM of 3–6 independent experiments using cells from different donors. Significant difference vs . unstimulated control: * P <0.05. Significant difference vs . NaCl control: • P <0.05.

Techniques: Cell Culture, Quantitative RT-PCR

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