vegfr 2 kinase activity  (Cell Signaling Technology Inc)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Name:
    HTScan VEGF Receptor 2 Kinase Assay Kit
    Description:
    The kit provides a means of performing kinase activity assays with recombinant human VEGFR 2 kinase It includes active VEGFR 2 kinase supplied as a GST fusion protein a biotinylated peptide substrate and a phospho tyrosine antibody for detection of the phosphorylated form of the substrate peptide
    Catalog Number:
    7788
    Price:
    None
    Category:
    Cellular Assay Kits
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc vegfr 2 kinase activity
    Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 <t>(VEGFR-2)</t> signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p
    The kit provides a means of performing kinase activity assays with recombinant human VEGFR 2 kinase It includes active VEGFR 2 kinase supplied as a GST fusion protein a biotinylated peptide substrate and a phospho tyrosine antibody for detection of the phosphorylated form of the substrate peptide
    https://www.bioz.com/result/vegfr 2 kinase activity/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vegfr 2 kinase activity - by Bioz Stars, 2021-04
    94/100 stars

    Images

    1) Product Images from "Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis"

    Article Title: Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis

    Journal: Journal of Breast Cancer

    doi: 10.4048/jbc.2018.21.1.11

    Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p
    Figure Legend Snippet: Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p

    Techniques Used: In Vivo, Multiple Displacement Amplification, Injection, Mouse Assay, Software, Immunohistochemistry, Staining, Expressing

    Isomangiferin suppresses breast cancer through inhibiting the vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway. (A) Isomangiferin induced cell apoptosis. Proteins from MDA-MB-231 cells treated with indicated concentrations of isomangiferin for 24 hours were submitted to Western blot for the immunoblotting of caspase-3 and cleaved poly ADP-ribose polymerase (cleaved PARP). (B) Isomangiferin's ihibiton on breast cancer cell proliferation was dependent on VEGFR-2 activity. 100 nM SU5408 was used or not to block VEGFR-2's activity and then MDA-MB-231 cells were treated with isomangiferin. The cell proliferation was assessed by MTS assay. Values are shown as mean±standard error of the mean of three independent experiments. (C) Isomangiferin suppressed the activation of VEGFR-2 in human umbilical vein endothelial cells (HUVECs). The activation of VEGFR-2 was analyzed by Western blot and probed with indicated antibodies. Western blot was conducted in the way that described in Methods and specific antibodies were used accordingly. PARP=poly ADP-ribose polymerase; NS=not significant; p-AKT=phosphorylated protein kinase B, p-PKB/p-AKT; AKT=protein kinase B, PKB/AKT; p-ERK=phosphorylated extracellular regulated protein kinases; ERK=extracellular regulated protein kinases; p-STAT3=phosphorylated signal transducer and activator of transcription 3; FAK=focal adhesion kinase. * p
    Figure Legend Snippet: Isomangiferin suppresses breast cancer through inhibiting the vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway. (A) Isomangiferin induced cell apoptosis. Proteins from MDA-MB-231 cells treated with indicated concentrations of isomangiferin for 24 hours were submitted to Western blot for the immunoblotting of caspase-3 and cleaved poly ADP-ribose polymerase (cleaved PARP). (B) Isomangiferin's ihibiton on breast cancer cell proliferation was dependent on VEGFR-2 activity. 100 nM SU5408 was used or not to block VEGFR-2's activity and then MDA-MB-231 cells were treated with isomangiferin. The cell proliferation was assessed by MTS assay. Values are shown as mean±standard error of the mean of three independent experiments. (C) Isomangiferin suppressed the activation of VEGFR-2 in human umbilical vein endothelial cells (HUVECs). The activation of VEGFR-2 was analyzed by Western blot and probed with indicated antibodies. Western blot was conducted in the way that described in Methods and specific antibodies were used accordingly. PARP=poly ADP-ribose polymerase; NS=not significant; p-AKT=phosphorylated protein kinase B, p-PKB/p-AKT; AKT=protein kinase B, PKB/AKT; p-ERK=phosphorylated extracellular regulated protein kinases; ERK=extracellular regulated protein kinases; p-STAT3=phosphorylated signal transducer and activator of transcription 3; FAK=focal adhesion kinase. * p

    Techniques Used: Multiple Displacement Amplification, Western Blot, Activity Assay, Blocking Assay, MTS Assay, Activation Assay

    2) Product Images from "Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis"

    Article Title: Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis

    Journal: Journal of Breast Cancer

    doi: 10.4048/jbc.2018.21.1.11

    Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p
    Figure Legend Snippet: Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p

    Techniques Used: In Vivo, Multiple Displacement Amplification, Injection, Mouse Assay, Software, Immunohistochemistry, Staining, Expressing

    Isomangiferin suppresses breast cancer through inhibiting the vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway. (A) Isomangiferin induced cell apoptosis. Proteins from MDA-MB-231 cells treated with indicated concentrations of isomangiferin for 24 hours were submitted to Western blot for the immunoblotting of caspase-3 and cleaved poly ADP-ribose polymerase (cleaved PARP). (B) Isomangiferin's ihibiton on breast cancer cell proliferation was dependent on VEGFR-2 activity. 100 nM SU5408 was used or not to block VEGFR-2's activity and then MDA-MB-231 cells were treated with isomangiferin. The cell proliferation was assessed by MTS assay. Values are shown as mean±standard error of the mean of three independent experiments. (C) Isomangiferin suppressed the activation of VEGFR-2 in human umbilical vein endothelial cells (HUVECs). The activation of VEGFR-2 was analyzed by Western blot and probed with indicated antibodies. Western blot was conducted in the way that described in Methods and specific antibodies were used accordingly. PARP=poly ADP-ribose polymerase; NS=not significant; p-AKT=phosphorylated protein kinase B, p-PKB/p-AKT; AKT=protein kinase B, PKB/AKT; p-ERK=phosphorylated extracellular regulated protein kinases; ERK=extracellular regulated protein kinases; p-STAT3=phosphorylated signal transducer and activator of transcription 3; FAK=focal adhesion kinase. * p
    Figure Legend Snippet: Isomangiferin suppresses breast cancer through inhibiting the vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway. (A) Isomangiferin induced cell apoptosis. Proteins from MDA-MB-231 cells treated with indicated concentrations of isomangiferin for 24 hours were submitted to Western blot for the immunoblotting of caspase-3 and cleaved poly ADP-ribose polymerase (cleaved PARP). (B) Isomangiferin's ihibiton on breast cancer cell proliferation was dependent on VEGFR-2 activity. 100 nM SU5408 was used or not to block VEGFR-2's activity and then MDA-MB-231 cells were treated with isomangiferin. The cell proliferation was assessed by MTS assay. Values are shown as mean±standard error of the mean of three independent experiments. (C) Isomangiferin suppressed the activation of VEGFR-2 in human umbilical vein endothelial cells (HUVECs). The activation of VEGFR-2 was analyzed by Western blot and probed with indicated antibodies. Western blot was conducted in the way that described in Methods and specific antibodies were used accordingly. PARP=poly ADP-ribose polymerase; NS=not significant; p-AKT=phosphorylated protein kinase B, p-PKB/p-AKT; AKT=protein kinase B, PKB/AKT; p-ERK=phosphorylated extracellular regulated protein kinases; ERK=extracellular regulated protein kinases; p-STAT3=phosphorylated signal transducer and activator of transcription 3; FAK=focal adhesion kinase. * p

    Techniques Used: Multiple Displacement Amplification, Western Blot, Activity Assay, Blocking Assay, MTS Assay, Activation Assay

    Related Articles

    Western Blot:

    Article Title: Acetyl-11-Keto-?-Boswellic Acid Inhibits Prostate Tumor Growth by Suppressing Vascular Endothelial Growth Factor Receptor 2-Mediated Angiogenesis
    Article Snippet: .. To confirm our Western blot analysis data and determine whether AKBA directly inhibits VEGFR2 kinase activity, we performed in vitro kinase assays with different concentrations of AKBA using HTScan® VEGF receptor 2 kinase assay kit according to manufacturer suggested methods (Cell Signaling Technology and PerkinElmer Life Sciences, USA). .. Our data demonstrated that AKBA directly inhibited VEGFR2 kinase activity in a dose-dependent manner with a 50% inhibitory concentration of 1.68 μMol/L ( ).

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: We found that 20 µM QODG dramatically inhibited phosphorylation of VEGFR2 Tyr1175 , but did not inhibit total VEGFR2 protein expression ( ). .. To confirm our Western blotting analysis data and verify whether QODG directly inhibits VEGFR2 tyrosine kinase activity, we performed an ELISA-based in vitro VEGFR2 tyrosine kinase assay with various concentrations of QODG using HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Massachusetts, USA) according to the manufacturer's suggested methods. .. As shown in , QODG directly inhibited VEGFR2 tyrosine kinase activity in a dose-dependent manner with a 50% inhibitory concentration (IC50 ) of 20.12 nM.

    Activity Assay:

    Article Title: Acetyl-11-Keto-?-Boswellic Acid Inhibits Prostate Tumor Growth by Suppressing Vascular Endothelial Growth Factor Receptor 2-Mediated Angiogenesis
    Article Snippet: .. To confirm our Western blot analysis data and determine whether AKBA directly inhibits VEGFR2 kinase activity, we performed in vitro kinase assays with different concentrations of AKBA using HTScan® VEGF receptor 2 kinase assay kit according to manufacturer suggested methods (Cell Signaling Technology and PerkinElmer Life Sciences, USA). .. Our data demonstrated that AKBA directly inhibited VEGFR2 kinase activity in a dose-dependent manner with a 50% inhibitory concentration of 1.68 μMol/L ( ).

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: .. In vitro VEGFR2 kinase inhibition assay In vitro the ability of QODG to inhibit VEGFR2 tyrosine kinase activity was assayed by prediluted QODG following the manual of HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Danvers, MA, USA). .. 4×reaction cocktail containing VEGFR2 was incubated with prediluted QODG or DMSO (0.1%) for 5 min at room temperature, and then 2×ATP/substrate peptide cocktail was added to the pre-incubated reaction cocktail/QODG compound or DMSO (0.1%).

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: We found that 20 µM QODG dramatically inhibited phosphorylation of VEGFR2 Tyr1175 , but did not inhibit total VEGFR2 protein expression ( ). .. To confirm our Western blotting analysis data and verify whether QODG directly inhibits VEGFR2 tyrosine kinase activity, we performed an ELISA-based in vitro VEGFR2 tyrosine kinase assay with various concentrations of QODG using HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Massachusetts, USA) according to the manufacturer's suggested methods. .. As shown in , QODG directly inhibited VEGFR2 tyrosine kinase activity in a dose-dependent manner with a 50% inhibitory concentration (IC50 ) of 20.12 nM.

    Article Title: Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis
    Article Snippet: After fixation with 4% paraformaldehyde and staining with 1% crystal violet, the number of adhering cells was quantified using the aforementioned inverted microscope. .. VEGFR-2 kinase activity (CST HTScan®, Cat No. #7788) was performed using a VEGFR-2 kinase assay kit purchased from Cell Signaling Technology. .. Briefly, the kinase activity was measured in a radiometric assay using the following reaction ingredients: 4 mM 3-(N-morpholino) propanesulfonic acid (pH 7.2), 2.5 mM β-glycerophosphate, 1 mM ethylene glycol-bis-(2-aminoethyl ether)-N,N′-tetraacetic acid, 0.4 mM ethylenediamine tetraacetic acid (EDTA), 4 mM magnesium chloride hexahydrate, 0.05 mM dithiothreitol, and 50 µM adenosine triphosphate.

    Article Title: Trans-3,4,5,4?-tetramethoxystilbene, a resveratrol analog, potently inhibits angiogenesis in vitro and in vivo
    Article Snippet: The reverse-transcribed RNA was primed with oligonucleotides specific for VEGFR1 (forward: 5′-GGGCAGACTCTTGTCCTCAACT-3′ reverse: 5′-CAGCTCATTTGCACCCTCGT-3′), VEGFR2 (forward: 5′-GACTGTGGCGAAGTGTTTTTGA-3′ reverse: 5′-GTGCAGGGGAGGGTTGGCGTAG-3′), and β-actin (forward: 5′-GTGCGGGACATCAAGGAGAA-3′ reverse: 5′-AGGAAGGAGGGCTGGAAGAG-3′) (Applied Biosystems, USA). .. In vitro VEGFR1 and VEGFR2 kinases activity assays After treatment, the tyrosine kinase activities of VEGFR1 and VEGFR2 were assayed with HTScan VEGFR1 and VEGFR2 kinase assay kits following the manufacturer's protocol (Cell Signaling Technology, USA). .. The results are expressed as percent kinase activity of the control (100%).

    In Vitro:

    Article Title: Acetyl-11-Keto-?-Boswellic Acid Inhibits Prostate Tumor Growth by Suppressing Vascular Endothelial Growth Factor Receptor 2-Mediated Angiogenesis
    Article Snippet: .. To confirm our Western blot analysis data and determine whether AKBA directly inhibits VEGFR2 kinase activity, we performed in vitro kinase assays with different concentrations of AKBA using HTScan® VEGF receptor 2 kinase assay kit according to manufacturer suggested methods (Cell Signaling Technology and PerkinElmer Life Sciences, USA). .. Our data demonstrated that AKBA directly inhibited VEGFR2 kinase activity in a dose-dependent manner with a 50% inhibitory concentration of 1.68 μMol/L ( ).

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: .. In vitro VEGFR2 kinase inhibition assay In vitro the ability of QODG to inhibit VEGFR2 tyrosine kinase activity was assayed by prediluted QODG following the manual of HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Danvers, MA, USA). .. 4×reaction cocktail containing VEGFR2 was incubated with prediluted QODG or DMSO (0.1%) for 5 min at room temperature, and then 2×ATP/substrate peptide cocktail was added to the pre-incubated reaction cocktail/QODG compound or DMSO (0.1%).

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: We found that 20 µM QODG dramatically inhibited phosphorylation of VEGFR2 Tyr1175 , but did not inhibit total VEGFR2 protein expression ( ). .. To confirm our Western blotting analysis data and verify whether QODG directly inhibits VEGFR2 tyrosine kinase activity, we performed an ELISA-based in vitro VEGFR2 tyrosine kinase assay with various concentrations of QODG using HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Massachusetts, USA) according to the manufacturer's suggested methods. .. As shown in , QODG directly inhibited VEGFR2 tyrosine kinase activity in a dose-dependent manner with a 50% inhibitory concentration (IC50 ) of 20.12 nM.

    Article Title: Trans-3,4,5,4?-tetramethoxystilbene, a resveratrol analog, potently inhibits angiogenesis in vitro and in vivo
    Article Snippet: The reverse-transcribed RNA was primed with oligonucleotides specific for VEGFR1 (forward: 5′-GGGCAGACTCTTGTCCTCAACT-3′ reverse: 5′-CAGCTCATTTGCACCCTCGT-3′), VEGFR2 (forward: 5′-GACTGTGGCGAAGTGTTTTTGA-3′ reverse: 5′-GTGCAGGGGAGGGTTGGCGTAG-3′), and β-actin (forward: 5′-GTGCGGGACATCAAGGAGAA-3′ reverse: 5′-AGGAAGGAGGGCTGGAAGAG-3′) (Applied Biosystems, USA). .. In vitro VEGFR1 and VEGFR2 kinases activity assays After treatment, the tyrosine kinase activities of VEGFR1 and VEGFR2 were assayed with HTScan VEGFR1 and VEGFR2 kinase assay kits following the manufacturer's protocol (Cell Signaling Technology, USA). .. The results are expressed as percent kinase activity of the control (100%).

    Article Title: α-santalol inhibits the angiogenesis and growth of human prostate tumor growth by targeting vascular endothelial growth factor receptor 2-mediated AKT/mTOR/P70S6K signaling pathway
    Article Snippet: .. In vitro VEGFR2 kinase inhibition assay VEGFR2 kinase assay was done using an HTScan® VEGFR2 kinase kit from Cell Signaling Technology (Cell Signaling Technology, Danvers, MA, USA) combined with colorimetric ELISA detection as described previously [ ]. .. The results were expressed as percent kinase activity of the vehicle control (100%), and IC50 was defined as the compound concentration that resulted in 50% inhibition of enzyme activity.

    Kinase Assay:

    Article Title: Acetyl-11-Keto-?-Boswellic Acid Inhibits Prostate Tumor Growth by Suppressing Vascular Endothelial Growth Factor Receptor 2-Mediated Angiogenesis
    Article Snippet: .. To confirm our Western blot analysis data and determine whether AKBA directly inhibits VEGFR2 kinase activity, we performed in vitro kinase assays with different concentrations of AKBA using HTScan® VEGF receptor 2 kinase assay kit according to manufacturer suggested methods (Cell Signaling Technology and PerkinElmer Life Sciences, USA). .. Our data demonstrated that AKBA directly inhibited VEGFR2 kinase activity in a dose-dependent manner with a 50% inhibitory concentration of 1.68 μMol/L ( ).

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: .. In vitro VEGFR2 kinase inhibition assay In vitro the ability of QODG to inhibit VEGFR2 tyrosine kinase activity was assayed by prediluted QODG following the manual of HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Danvers, MA, USA). .. 4×reaction cocktail containing VEGFR2 was incubated with prediluted QODG or DMSO (0.1%) for 5 min at room temperature, and then 2×ATP/substrate peptide cocktail was added to the pre-incubated reaction cocktail/QODG compound or DMSO (0.1%).

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: We found that 20 µM QODG dramatically inhibited phosphorylation of VEGFR2 Tyr1175 , but did not inhibit total VEGFR2 protein expression ( ). .. To confirm our Western blotting analysis data and verify whether QODG directly inhibits VEGFR2 tyrosine kinase activity, we performed an ELISA-based in vitro VEGFR2 tyrosine kinase assay with various concentrations of QODG using HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Massachusetts, USA) according to the manufacturer's suggested methods. .. As shown in , QODG directly inhibited VEGFR2 tyrosine kinase activity in a dose-dependent manner with a 50% inhibitory concentration (IC50 ) of 20.12 nM.

    Article Title: Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis
    Article Snippet: After fixation with 4% paraformaldehyde and staining with 1% crystal violet, the number of adhering cells was quantified using the aforementioned inverted microscope. .. VEGFR-2 kinase activity (CST HTScan®, Cat No. #7788) was performed using a VEGFR-2 kinase assay kit purchased from Cell Signaling Technology. .. Briefly, the kinase activity was measured in a radiometric assay using the following reaction ingredients: 4 mM 3-(N-morpholino) propanesulfonic acid (pH 7.2), 2.5 mM β-glycerophosphate, 1 mM ethylene glycol-bis-(2-aminoethyl ether)-N,N′-tetraacetic acid, 0.4 mM ethylenediamine tetraacetic acid (EDTA), 4 mM magnesium chloride hexahydrate, 0.05 mM dithiothreitol, and 50 µM adenosine triphosphate.

    Article Title: Indirubin derivative E804 inhibits angiogenesis
    Article Snippet: After seven days, the microvessel growth was measured by taking photographs with the AxioImager ZI inverted microscope (Carl Zeiss) using a 4x objective lens. .. VEGFR-2 inhibition assay A 12.5 μL aliquot of the 4x reaction cocktail containing 100 ng VEGFR-2 [supplied from the HTScan VEGFR-2 kinase assay kit (Cell Signaling Technology)] was incubated with 12.5 μL of IDR-E804 for 5 min at room temperature. ..

    Article Title: Trans-3,4,5,4?-tetramethoxystilbene, a resveratrol analog, potently inhibits angiogenesis in vitro and in vivo
    Article Snippet: The reverse-transcribed RNA was primed with oligonucleotides specific for VEGFR1 (forward: 5′-GGGCAGACTCTTGTCCTCAACT-3′ reverse: 5′-CAGCTCATTTGCACCCTCGT-3′), VEGFR2 (forward: 5′-GACTGTGGCGAAGTGTTTTTGA-3′ reverse: 5′-GTGCAGGGGAGGGTTGGCGTAG-3′), and β-actin (forward: 5′-GTGCGGGACATCAAGGAGAA-3′ reverse: 5′-AGGAAGGAGGGCTGGAAGAG-3′) (Applied Biosystems, USA). .. In vitro VEGFR1 and VEGFR2 kinases activity assays After treatment, the tyrosine kinase activities of VEGFR1 and VEGFR2 were assayed with HTScan VEGFR1 and VEGFR2 kinase assay kits following the manufacturer's protocol (Cell Signaling Technology, USA). .. The results are expressed as percent kinase activity of the control (100%).

    Article Title: α-santalol inhibits the angiogenesis and growth of human prostate tumor growth by targeting vascular endothelial growth factor receptor 2-mediated AKT/mTOR/P70S6K signaling pathway
    Article Snippet: .. In vitro VEGFR2 kinase inhibition assay VEGFR2 kinase assay was done using an HTScan® VEGFR2 kinase kit from Cell Signaling Technology (Cell Signaling Technology, Danvers, MA, USA) combined with colorimetric ELISA detection as described previously [ ]. .. The results were expressed as percent kinase activity of the vehicle control (100%), and IC50 was defined as the compound concentration that resulted in 50% inhibition of enzyme activity.

    Inhibition:

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: .. In vitro VEGFR2 kinase inhibition assay In vitro the ability of QODG to inhibit VEGFR2 tyrosine kinase activity was assayed by prediluted QODG following the manual of HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Danvers, MA, USA). .. 4×reaction cocktail containing VEGFR2 was incubated with prediluted QODG or DMSO (0.1%) for 5 min at room temperature, and then 2×ATP/substrate peptide cocktail was added to the pre-incubated reaction cocktail/QODG compound or DMSO (0.1%).

    Article Title: Indirubin derivative E804 inhibits angiogenesis
    Article Snippet: After seven days, the microvessel growth was measured by taking photographs with the AxioImager ZI inverted microscope (Carl Zeiss) using a 4x objective lens. .. VEGFR-2 inhibition assay A 12.5 μL aliquot of the 4x reaction cocktail containing 100 ng VEGFR-2 [supplied from the HTScan VEGFR-2 kinase assay kit (Cell Signaling Technology)] was incubated with 12.5 μL of IDR-E804 for 5 min at room temperature. ..

    Article Title: α-santalol inhibits the angiogenesis and growth of human prostate tumor growth by targeting vascular endothelial growth factor receptor 2-mediated AKT/mTOR/P70S6K signaling pathway
    Article Snippet: .. In vitro VEGFR2 kinase inhibition assay VEGFR2 kinase assay was done using an HTScan® VEGFR2 kinase kit from Cell Signaling Technology (Cell Signaling Technology, Danvers, MA, USA) combined with colorimetric ELISA detection as described previously [ ]. .. The results were expressed as percent kinase activity of the vehicle control (100%), and IC50 was defined as the compound concentration that resulted in 50% inhibition of enzyme activity.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: We found that 20 µM QODG dramatically inhibited phosphorylation of VEGFR2 Tyr1175 , but did not inhibit total VEGFR2 protein expression ( ). .. To confirm our Western blotting analysis data and verify whether QODG directly inhibits VEGFR2 tyrosine kinase activity, we performed an ELISA-based in vitro VEGFR2 tyrosine kinase assay with various concentrations of QODG using HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Massachusetts, USA) according to the manufacturer's suggested methods. .. As shown in , QODG directly inhibited VEGFR2 tyrosine kinase activity in a dose-dependent manner with a 50% inhibitory concentration (IC50 ) of 20.12 nM.

    Article Title: α-santalol inhibits the angiogenesis and growth of human prostate tumor growth by targeting vascular endothelial growth factor receptor 2-mediated AKT/mTOR/P70S6K signaling pathway
    Article Snippet: .. In vitro VEGFR2 kinase inhibition assay VEGFR2 kinase assay was done using an HTScan® VEGFR2 kinase kit from Cell Signaling Technology (Cell Signaling Technology, Danvers, MA, USA) combined with colorimetric ELISA detection as described previously [ ]. .. The results were expressed as percent kinase activity of the vehicle control (100%), and IC50 was defined as the compound concentration that resulted in 50% inhibition of enzyme activity.

    Tyrosine Kinase Assay:

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells
    Article Snippet: We found that 20 µM QODG dramatically inhibited phosphorylation of VEGFR2 Tyr1175 , but did not inhibit total VEGFR2 protein expression ( ). .. To confirm our Western blotting analysis data and verify whether QODG directly inhibits VEGFR2 tyrosine kinase activity, we performed an ELISA-based in vitro VEGFR2 tyrosine kinase assay with various concentrations of QODG using HTScan® VEGFR2 kinase assay kit (Cell Signaling Technology, Massachusetts, USA) according to the manufacturer's suggested methods. .. As shown in , QODG directly inhibited VEGFR2 tyrosine kinase activity in a dose-dependent manner with a 50% inhibitory concentration (IC50 ) of 20.12 nM.

    Incubation:

    Article Title: Indirubin derivative E804 inhibits angiogenesis
    Article Snippet: After seven days, the microvessel growth was measured by taking photographs with the AxioImager ZI inverted microscope (Carl Zeiss) using a 4x objective lens. .. VEGFR-2 inhibition assay A 12.5 μL aliquot of the 4x reaction cocktail containing 100 ng VEGFR-2 [supplied from the HTScan VEGFR-2 kinase assay kit (Cell Signaling Technology)] was incubated with 12.5 μL of IDR-E804 for 5 min at room temperature. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Cell Signaling Technology Inc vegfr 2 phosphorylation
    Hydroxytyrosol inhibits vascular endothelial growth factor (VEGF)-induced <t>VEGFR-2</t> phosphorylation by interacting with cell membrane components. Human umbilical vein endothelial cells (HUVECs) were incubated with 50 μM or 1 μM hydroxytyrosol and VEGF (25 ng/mL). ( A ) Hydroxytyrosol and VEGF were mixed 5 min prior to cells treatment for another 5 min; ( B ) hydroxytyrosol was pre-incubated for 4 h with HUVECs and subsequently incubated with VEGF for 5 min; ( C ) hydroxytyrosol was pre-incubated for 4 h with HUVECs and, after removal of the polyphenol (with PBS), was then stimulated with VEGF for 5 min. p-VEGFR-2 (Tyr1175) was quantified by ELISA essay; * p
    Vegfr 2 Phosphorylation, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vegfr 2 phosphorylation/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vegfr 2 phosphorylation - by Bioz Stars, 2021-04
    99/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc vegfr 2 kinase activity
    Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 <t>(VEGFR-2)</t> signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p
    Vegfr 2 Kinase Activity, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vegfr 2 kinase activity/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vegfr 2 kinase activity - by Bioz Stars, 2021-04
    94/100 stars
      Buy from Supplier

    Image Search Results


    Hydroxytyrosol inhibits vascular endothelial growth factor (VEGF)-induced VEGFR-2 phosphorylation by interacting with cell membrane components. Human umbilical vein endothelial cells (HUVECs) were incubated with 50 μM or 1 μM hydroxytyrosol and VEGF (25 ng/mL). ( A ) Hydroxytyrosol and VEGF were mixed 5 min prior to cells treatment for another 5 min; ( B ) hydroxytyrosol was pre-incubated for 4 h with HUVECs and subsequently incubated with VEGF for 5 min; ( C ) hydroxytyrosol was pre-incubated for 4 h with HUVECs and, after removal of the polyphenol (with PBS), was then stimulated with VEGF for 5 min. p-VEGFR-2 (Tyr1175) was quantified by ELISA essay; * p

    Journal: Nutrients

    Article Title: Anti-VEGF Signalling Mechanism in HUVECs by Melatonin, Serotonin, Hydroxytyrosol and Other Bioactive Compounds

    doi: 10.3390/nu11102421

    Figure Lengend Snippet: Hydroxytyrosol inhibits vascular endothelial growth factor (VEGF)-induced VEGFR-2 phosphorylation by interacting with cell membrane components. Human umbilical vein endothelial cells (HUVECs) were incubated with 50 μM or 1 μM hydroxytyrosol and VEGF (25 ng/mL). ( A ) Hydroxytyrosol and VEGF were mixed 5 min prior to cells treatment for another 5 min; ( B ) hydroxytyrosol was pre-incubated for 4 h with HUVECs and subsequently incubated with VEGF for 5 min; ( C ) hydroxytyrosol was pre-incubated for 4 h with HUVECs and, after removal of the polyphenol (with PBS), was then stimulated with VEGF for 5 min. p-VEGFR-2 (Tyr1175) was quantified by ELISA essay; * p

    Article Snippet: VEGFR-2 Phosphorylation (ELISA Assay) The PathScan Phospho-VEGFR-2 (Tyr 1175) sandwich ELISA kit (Cell Signaling Technology, Hitchin, UK) was used to quantify VEGFR-2 activation in protein lysates, the procedure being performed following the manufacturer’s instructions.

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay

    VEGFA induces PC3 cell migration via VEGFR-2 activation. PC3 cells (3×10 5 per transwell insert) were ( a ) treated with different concentrations of recombinant VEGFA 165 for 5 h or ( b ) pretreated with the VEGFR-2 kinase inhibitor Ki8751 for 30 min

    Journal: Asian Journal of Andrology

    Article Title: Vascular endothelial growth factor A, secreted in response to transforming growth factor-?1 under hypoxic conditions, induces autocrine effects on migration of prostate cancer cells

    doi: 10.1038/aja.2011.197

    Figure Lengend Snippet: VEGFA induces PC3 cell migration via VEGFR-2 activation. PC3 cells (3×10 5 per transwell insert) were ( a ) treated with different concentrations of recombinant VEGFA 165 for 5 h or ( b ) pretreated with the VEGFR-2 kinase inhibitor Ki8751 for 30 min

    Article Snippet: VEGFR-2 was detected with anti-VEGFR-2 antibody (1∶1000; Cell Signaling, Danvers, MA, USA).

    Techniques: Migration, Activation Assay, Recombinant

    A diagram illustrating how hypoxia and TGF-β1 become tumorigenic in advanced prostate cancer cells to increase tumor metastasis. PC3 prostate cancer cell line expresses VEGFR-2 protein that enables an autocrine VEGFA signaling in PC3 cells. The

    Journal: Asian Journal of Andrology

    Article Title: Vascular endothelial growth factor A, secreted in response to transforming growth factor-?1 under hypoxic conditions, induces autocrine effects on migration of prostate cancer cells

    doi: 10.1038/aja.2011.197

    Figure Lengend Snippet: A diagram illustrating how hypoxia and TGF-β1 become tumorigenic in advanced prostate cancer cells to increase tumor metastasis. PC3 prostate cancer cell line expresses VEGFR-2 protein that enables an autocrine VEGFA signaling in PC3 cells. The

    Article Snippet: VEGFR-2 was detected with anti-VEGFR-2 antibody (1∶1000; Cell Signaling, Danvers, MA, USA).

    Techniques:

    Calcium dobesilate (CaD) inhibits VEGF-induced VEGFR-2 activation. VEGF 165 (25 ng/mL) was premixed and incubated with: ( A ) various CaD concentrations (6–100 μM) or ( B ) 50–100 μM CaD for 1 h, before exposure to HUVECs for 2 min. ( C ) HUVECs were incubated with 50 and 100μM CaD for 60 min with three subsequent washing steps with warm medium before stimulation with VEGF 165 for 2 min. Western blot analysis was performed using anti-phospho-VEGFR-2 antibody and total VEGFR2 was used as a loading control after membrane stripping. Each bar represents the mean ± SD (n = 3). *P

    Journal: PLoS ONE

    Article Title: Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice

    doi: 10.1371/journal.pone.0218494

    Figure Lengend Snippet: Calcium dobesilate (CaD) inhibits VEGF-induced VEGFR-2 activation. VEGF 165 (25 ng/mL) was premixed and incubated with: ( A ) various CaD concentrations (6–100 μM) or ( B ) 50–100 μM CaD for 1 h, before exposure to HUVECs for 2 min. ( C ) HUVECs were incubated with 50 and 100μM CaD for 60 min with three subsequent washing steps with warm medium before stimulation with VEGF 165 for 2 min. Western blot analysis was performed using anti-phospho-VEGFR-2 antibody and total VEGFR2 was used as a loading control after membrane stripping. Each bar represents the mean ± SD (n = 3). *P

    Article Snippet: Rabbit primary antibodies for VEGFR-2, p-Tyr1175, p-ERK1/2, ERK1/2, p-P38, p-MEK and MEK were acquired from Cell Signaling Technology (Leiden, The Netherlands) and F4/80 (clone A3-1; BioLegend, San Diego, CA, USA).

    Techniques: Activation Assay, Incubation, Western Blot, Stripping Membranes

    Postulated model of interactions between VEGF 165 , VEGFR-2, and CaD. VEGF 165 binds to its co-receptor heparin sulfates (HS) of the endothelial glycocalyx with a specific binding site (left box) which stabilizes the VEGF-VEGF-R binding leading to phosphorylation of VEGFR-2 receptor, intracellular signaling and cell activation (middle box). CaD interacts with the heparin-binding domain of the VEGF 165 (right box), thereby displacing HS from its binding site, and decreases VEGF-induced intracellular signaling. CaD also regulates VEGF 165 activity by participating in the formation of unstable VEGF-VEGFR-2 complex.

    Journal: PLoS ONE

    Article Title: Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice

    doi: 10.1371/journal.pone.0218494

    Figure Lengend Snippet: Postulated model of interactions between VEGF 165 , VEGFR-2, and CaD. VEGF 165 binds to its co-receptor heparin sulfates (HS) of the endothelial glycocalyx with a specific binding site (left box) which stabilizes the VEGF-VEGF-R binding leading to phosphorylation of VEGFR-2 receptor, intracellular signaling and cell activation (middle box). CaD interacts with the heparin-binding domain of the VEGF 165 (right box), thereby displacing HS from its binding site, and decreases VEGF-induced intracellular signaling. CaD also regulates VEGF 165 activity by participating in the formation of unstable VEGF-VEGFR-2 complex.

    Article Snippet: Rabbit primary antibodies for VEGFR-2, p-Tyr1175, p-ERK1/2, ERK1/2, p-P38, p-MEK and MEK were acquired from Cell Signaling Technology (Leiden, The Netherlands) and F4/80 (clone A3-1; BioLegend, San Diego, CA, USA).

    Techniques: Binding Assay, Activation Assay, Activity Assay

    CaD inhibits VEGFR-2 phosphorylation and signaling in STZ diabetic mice. STZ mice were daily treated with vehicle, CaD or Enalapril and sacrificed at week 12 for the analysis. Non diabetic mice were used as control. Kidneys were isolated and homogenized as described in materials and methods. Phosphorylated VEGFR-2 (A) in the kidney lysates was determined by ELISA mean ± SD of 5–6 animals. Phosphorylated ERK1/2 (B) was analyzed by Western blot as described in Fig 1 . Vinculin was used as a loading control. Phosphorylated P38 (C) was analyzed by immunohistochemistry as described in the materials and methods section. Autofluorescence is shown in green, scale bar 100 μM. Representative image for n = 6/condition is depicted. *P

    Journal: PLoS ONE

    Article Title: Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice

    doi: 10.1371/journal.pone.0218494

    Figure Lengend Snippet: CaD inhibits VEGFR-2 phosphorylation and signaling in STZ diabetic mice. STZ mice were daily treated with vehicle, CaD or Enalapril and sacrificed at week 12 for the analysis. Non diabetic mice were used as control. Kidneys were isolated and homogenized as described in materials and methods. Phosphorylated VEGFR-2 (A) in the kidney lysates was determined by ELISA mean ± SD of 5–6 animals. Phosphorylated ERK1/2 (B) was analyzed by Western blot as described in Fig 1 . Vinculin was used as a loading control. Phosphorylated P38 (C) was analyzed by immunohistochemistry as described in the materials and methods section. Autofluorescence is shown in green, scale bar 100 μM. Representative image for n = 6/condition is depicted. *P

    Article Snippet: Rabbit primary antibodies for VEGFR-2, p-Tyr1175, p-ERK1/2, ERK1/2, p-P38, p-MEK and MEK were acquired from Cell Signaling Technology (Leiden, The Netherlands) and F4/80 (clone A3-1; BioLegend, San Diego, CA, USA).

    Techniques: Mouse Assay, Isolation, Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemistry

    Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p

    Journal: Journal of Breast Cancer

    Article Title: Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis

    doi: 10.4048/jbc.2018.21.1.11

    Figure Lengend Snippet: Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p

    Article Snippet: VEGFR-2 kinase activity (CST HTScan®, Cat No. #7788) was performed using a VEGFR-2 kinase assay kit purchased from Cell Signaling Technology.

    Techniques: In Vivo, Multiple Displacement Amplification, Injection, Mouse Assay, Software, Immunohistochemistry, Staining, Expressing

    Isomangiferin suppresses breast cancer through inhibiting the vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway. (A) Isomangiferin induced cell apoptosis. Proteins from MDA-MB-231 cells treated with indicated concentrations of isomangiferin for 24 hours were submitted to Western blot for the immunoblotting of caspase-3 and cleaved poly ADP-ribose polymerase (cleaved PARP). (B) Isomangiferin's ihibiton on breast cancer cell proliferation was dependent on VEGFR-2 activity. 100 nM SU5408 was used or not to block VEGFR-2's activity and then MDA-MB-231 cells were treated with isomangiferin. The cell proliferation was assessed by MTS assay. Values are shown as mean±standard error of the mean of three independent experiments. (C) Isomangiferin suppressed the activation of VEGFR-2 in human umbilical vein endothelial cells (HUVECs). The activation of VEGFR-2 was analyzed by Western blot and probed with indicated antibodies. Western blot was conducted in the way that described in Methods and specific antibodies were used accordingly. PARP=poly ADP-ribose polymerase; NS=not significant; p-AKT=phosphorylated protein kinase B, p-PKB/p-AKT; AKT=protein kinase B, PKB/AKT; p-ERK=phosphorylated extracellular regulated protein kinases; ERK=extracellular regulated protein kinases; p-STAT3=phosphorylated signal transducer and activator of transcription 3; FAK=focal adhesion kinase. * p

    Journal: Journal of Breast Cancer

    Article Title: Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis

    doi: 10.4048/jbc.2018.21.1.11

    Figure Lengend Snippet: Isomangiferin suppresses breast cancer through inhibiting the vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway. (A) Isomangiferin induced cell apoptosis. Proteins from MDA-MB-231 cells treated with indicated concentrations of isomangiferin for 24 hours were submitted to Western blot for the immunoblotting of caspase-3 and cleaved poly ADP-ribose polymerase (cleaved PARP). (B) Isomangiferin's ihibiton on breast cancer cell proliferation was dependent on VEGFR-2 activity. 100 nM SU5408 was used or not to block VEGFR-2's activity and then MDA-MB-231 cells were treated with isomangiferin. The cell proliferation was assessed by MTS assay. Values are shown as mean±standard error of the mean of three independent experiments. (C) Isomangiferin suppressed the activation of VEGFR-2 in human umbilical vein endothelial cells (HUVECs). The activation of VEGFR-2 was analyzed by Western blot and probed with indicated antibodies. Western blot was conducted in the way that described in Methods and specific antibodies were used accordingly. PARP=poly ADP-ribose polymerase; NS=not significant; p-AKT=phosphorylated protein kinase B, p-PKB/p-AKT; AKT=protein kinase B, PKB/AKT; p-ERK=phosphorylated extracellular regulated protein kinases; ERK=extracellular regulated protein kinases; p-STAT3=phosphorylated signal transducer and activator of transcription 3; FAK=focal adhesion kinase. * p

    Article Snippet: VEGFR-2 kinase activity (CST HTScan®, Cat No. #7788) was performed using a VEGFR-2 kinase assay kit purchased from Cell Signaling Technology.

    Techniques: Multiple Displacement Amplification, Western Blot, Activity Assay, Blocking Assay, MTS Assay, Activation Assay