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  • 99
    Millipore sodium orthovanada
    Treatment with <t>EGF</t> does not affect phosphorylation of EGFRvIII ( A ) DK-MG cells treated with indicated concentration of EGF for 10 min were lysed and analysed by Western blotting. ( B ) Quantification of EGFRvIII phosphorylation as shown in A. Statistical analysis performed using one-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. ( C ) Western blot analysis of cells stimulated for 1 h with 20 ng/mL EGF, 1 mM <t>NaOVa,</t> concomitant EGF and NaOVa or Control cells, as indicated. ( D ) Quantification of blots as shown in C, with normalization to wild-type receptor under Control conditions. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. *** p
    Sodium Orthovanada, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 147 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Price from $9.99 to $1999.99
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    90
    Selleck Chemicals sodium orthovanadate
    Treatment with <t>EGF</t> does not affect phosphorylation of EGFRvIII ( A ) DK-MG cells treated with indicated concentration of EGF for 10 min were lysed and analysed by Western blotting. ( B ) Quantification of EGFRvIII phosphorylation as shown in A. Statistical analysis performed using one-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. ( C ) Western blot analysis of cells stimulated for 1 h with 20 ng/mL EGF, 1 mM <t>NaOVa,</t> concomitant EGF and NaOVa or Control cells, as indicated. ( D ) Quantification of blots as shown in C, with normalization to wild-type receptor under Control conditions. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. *** p
    Sodium Orthovanadate, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sodium orthovanadate/product/Selleck Chemicals
    Average 90 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
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    85
    Tocris sodium orthovanadate
    Treatment with <t>EGF</t> does not affect phosphorylation of EGFRvIII ( A ) DK-MG cells treated with indicated concentration of EGF for 10 min were lysed and analysed by Western blotting. ( B ) Quantification of EGFRvIII phosphorylation as shown in A. Statistical analysis performed using one-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. ( C ) Western blot analysis of cells stimulated for 1 h with 20 ng/mL EGF, 1 mM <t>NaOVa,</t> concomitant EGF and NaOVa or Control cells, as indicated. ( D ) Quantification of blots as shown in C, with normalization to wild-type receptor under Control conditions. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. *** p
    Sodium Orthovanadate, supplied by Tocris, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sodium orthovanadate/product/Tocris
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    Image Search Results


    Treatment with EGF does not affect phosphorylation of EGFRvIII ( A ) DK-MG cells treated with indicated concentration of EGF for 10 min were lysed and analysed by Western blotting. ( B ) Quantification of EGFRvIII phosphorylation as shown in A. Statistical analysis performed using one-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. ( C ) Western blot analysis of cells stimulated for 1 h with 20 ng/mL EGF, 1 mM NaOVa, concomitant EGF and NaOVa or Control cells, as indicated. ( D ) Quantification of blots as shown in C, with normalization to wild-type receptor under Control conditions. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. *** p

    Journal: Oncotarget

    Article Title: Cyclic trans-phosphorylation in a homodimer as the predominant mechanism of EGFRvIII action and regulation

    doi: 10.18632/oncotarget.24058

    Figure Lengend Snippet: Treatment with EGF does not affect phosphorylation of EGFRvIII ( A ) DK-MG cells treated with indicated concentration of EGF for 10 min were lysed and analysed by Western blotting. ( B ) Quantification of EGFRvIII phosphorylation as shown in A. Statistical analysis performed using one-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. ( C ) Western blot analysis of cells stimulated for 1 h with 20 ng/mL EGF, 1 mM NaOVa, concomitant EGF and NaOVa or Control cells, as indicated. ( D ) Quantification of blots as shown in C, with normalization to wild-type receptor under Control conditions. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. *** p

    Article Snippet: Chemicals To analyse the effects of pharmacological inhibitors, cells were seeded in 6-well plates at 2.5x105 cells per well, incubated overnight to allow for adhesion and serum starved for subsequent 24 h. Serum free medium containing appropriate inhibitors was used to pre-treat cells for 30 min, unless indicated otherwise, prior to stimulation with 20 ng/mL EGF (Sigma-Aldrich, St Louis, MO, USA, Cat. no. E5036), 1 mM sodium orthovanadate – NaOVa (Na3 VO4 , Calbiochem, Cat. no. 567540), phenylarsine oxide – PAO (Sigma, Cat. no. P3075) or pervanadate – pV (obtained as described previously [ ]; using Catalase, Sigma, Cat. no. C1345 and H2 O2 Sigma, Cat. no. 31642) for 1 hour and lysed.

    Techniques: Concentration Assay, Western Blot

    Homodimerization of EGFRvIII ( A ) Semi-native Western blot (non-reducing conditions) analysis of the AD293 cell lines expressing EGFRvIII or EGFRvIIIC16S treated simultaneously with EGFR TKIs and NaOVa, as indicated. Arrowheads indicate dimers, arrows point to monomers. ( B ) DK-MG low , cell line with marginal endogenous EGFRvIII expression, were transduced to constitutively express either naïve EGFRvIII (DK-MG exo ) or C16S mutated version (DK-MG C16S ). Cells were treated with NaOVa or EGF, as indicated. Immunoblots have been uniformly adjusted for brightness and contrast to facilitate interpretation.

    Journal: Oncotarget

    Article Title: Cyclic trans-phosphorylation in a homodimer as the predominant mechanism of EGFRvIII action and regulation

    doi: 10.18632/oncotarget.24058

    Figure Lengend Snippet: Homodimerization of EGFRvIII ( A ) Semi-native Western blot (non-reducing conditions) analysis of the AD293 cell lines expressing EGFRvIII or EGFRvIIIC16S treated simultaneously with EGFR TKIs and NaOVa, as indicated. Arrowheads indicate dimers, arrows point to monomers. ( B ) DK-MG low , cell line with marginal endogenous EGFRvIII expression, were transduced to constitutively express either naïve EGFRvIII (DK-MG exo ) or C16S mutated version (DK-MG C16S ). Cells were treated with NaOVa or EGF, as indicated. Immunoblots have been uniformly adjusted for brightness and contrast to facilitate interpretation.

    Article Snippet: Chemicals To analyse the effects of pharmacological inhibitors, cells were seeded in 6-well plates at 2.5x105 cells per well, incubated overnight to allow for adhesion and serum starved for subsequent 24 h. Serum free medium containing appropriate inhibitors was used to pre-treat cells for 30 min, unless indicated otherwise, prior to stimulation with 20 ng/mL EGF (Sigma-Aldrich, St Louis, MO, USA, Cat. no. E5036), 1 mM sodium orthovanadate – NaOVa (Na3 VO4 , Calbiochem, Cat. no. 567540), phenylarsine oxide – PAO (Sigma, Cat. no. P3075) or pervanadate – pV (obtained as described previously [ ]; using Catalase, Sigma, Cat. no. C1345 and H2 O2 Sigma, Cat. no. 31642) for 1 hour and lysed.

    Techniques: Western Blot, Expressing

    Treatment of DK-MG cells with phosphatase inhibitors results in hyperphosphorylation of EGFRwt and EGFRvIII on majority of tyrosine residues ( A ) Western blot analysis of EGFR phosphorylation in cells treated with NaOVa, pV or PAO at indicated concentrations for 1 h. ( B ) Analysis of phosphorylation status of EGFR on selected tyrosine residues following stimulation with 20 ng/mL EGF, 1 mM NaOVa or 0.5 µM PAO. ( C–F ) Ratio of phosphorylated tyrosine to the total EGFR protein for the wild-type and mutant protein following stimulation as indicated in B, for residue Tyr 1045 (C), Tyr1068 (D), Tyr1148 (E) and Tyr 1173 (F). Statistical analysis performed using one-way ANOVA with post-analysis Bonferroni’s multiple comparisons test for each residue within one receptor variant. ( G ) Quantification of phosphorylated tyrosine 1068 to total EGFR for EGFRwt and EGFRvIII is presented. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test, n = 23. *** p

    Journal: Oncotarget

    Article Title: Cyclic trans-phosphorylation in a homodimer as the predominant mechanism of EGFRvIII action and regulation

    doi: 10.18632/oncotarget.24058

    Figure Lengend Snippet: Treatment of DK-MG cells with phosphatase inhibitors results in hyperphosphorylation of EGFRwt and EGFRvIII on majority of tyrosine residues ( A ) Western blot analysis of EGFR phosphorylation in cells treated with NaOVa, pV or PAO at indicated concentrations for 1 h. ( B ) Analysis of phosphorylation status of EGFR on selected tyrosine residues following stimulation with 20 ng/mL EGF, 1 mM NaOVa or 0.5 µM PAO. ( C–F ) Ratio of phosphorylated tyrosine to the total EGFR protein for the wild-type and mutant protein following stimulation as indicated in B, for residue Tyr 1045 (C), Tyr1068 (D), Tyr1148 (E) and Tyr 1173 (F). Statistical analysis performed using one-way ANOVA with post-analysis Bonferroni’s multiple comparisons test for each residue within one receptor variant. ( G ) Quantification of phosphorylated tyrosine 1068 to total EGFR for EGFRwt and EGFRvIII is presented. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test, n = 23. *** p

    Article Snippet: Chemicals To analyse the effects of pharmacological inhibitors, cells were seeded in 6-well plates at 2.5x105 cells per well, incubated overnight to allow for adhesion and serum starved for subsequent 24 h. Serum free medium containing appropriate inhibitors was used to pre-treat cells for 30 min, unless indicated otherwise, prior to stimulation with 20 ng/mL EGF (Sigma-Aldrich, St Louis, MO, USA, Cat. no. E5036), 1 mM sodium orthovanadate – NaOVa (Na3 VO4 , Calbiochem, Cat. no. 567540), phenylarsine oxide – PAO (Sigma, Cat. no. P3075) or pervanadate – pV (obtained as described previously [ ]; using Catalase, Sigma, Cat. no. C1345 and H2 O2 Sigma, Cat. no. 31642) for 1 hour and lysed.

    Techniques: Western Blot, Mutagenesis, Variant Assay

    EGFRvIII is not phosphorylated by EGFRwt ( A ) Western blot analysis of cells treated with indicated inhibitors prior to and concurrently with EGF or NaOVa treatment for 1 h. Concentrations of inhibitors were based on literature reports. ( B ) Cells were treated with inhibitors specific to pan-JAK (ruxolitinib), CDK2 (flavopiridol) or DNA-PK (Nu-7441) prior to and concurrently with NaOVa treatment. Concentrations of inhibitors were based on literature reports. ( C ) Cells transiently expressing control scrambled (SCR) shRNA or shRNA targeted against EGFRwt were treated as indicated and analyzed by Western blotting. ( D ) Quantification of blots as shown in C, with columns representing ratio of phosphorylated to total EGFRvIII protein. ( E ) Cells treated with cycloheximide and stimulated with EGF for 3 h as indicated. Where indicated, cells were incubated for another 30 min with DMSO or gefitinib prior to 1 h treatment with NaOVa. ( F ) Quantification of blots as in E for EGFRvIII, with normalization to DMSO treated Control cells. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. ** p

    Journal: Oncotarget

    Article Title: Cyclic trans-phosphorylation in a homodimer as the predominant mechanism of EGFRvIII action and regulation

    doi: 10.18632/oncotarget.24058

    Figure Lengend Snippet: EGFRvIII is not phosphorylated by EGFRwt ( A ) Western blot analysis of cells treated with indicated inhibitors prior to and concurrently with EGF or NaOVa treatment for 1 h. Concentrations of inhibitors were based on literature reports. ( B ) Cells were treated with inhibitors specific to pan-JAK (ruxolitinib), CDK2 (flavopiridol) or DNA-PK (Nu-7441) prior to and concurrently with NaOVa treatment. Concentrations of inhibitors were based on literature reports. ( C ) Cells transiently expressing control scrambled (SCR) shRNA or shRNA targeted against EGFRwt were treated as indicated and analyzed by Western blotting. ( D ) Quantification of blots as shown in C, with columns representing ratio of phosphorylated to total EGFRvIII protein. ( E ) Cells treated with cycloheximide and stimulated with EGF for 3 h as indicated. Where indicated, cells were incubated for another 30 min with DMSO or gefitinib prior to 1 h treatment with NaOVa. ( F ) Quantification of blots as in E for EGFRvIII, with normalization to DMSO treated Control cells. Statistical analysis performed using two-way ANOVA with post-analysis Bonferroni’s multiple comparisons test. ** p

    Article Snippet: Chemicals To analyse the effects of pharmacological inhibitors, cells were seeded in 6-well plates at 2.5x105 cells per well, incubated overnight to allow for adhesion and serum starved for subsequent 24 h. Serum free medium containing appropriate inhibitors was used to pre-treat cells for 30 min, unless indicated otherwise, prior to stimulation with 20 ng/mL EGF (Sigma-Aldrich, St Louis, MO, USA, Cat. no. E5036), 1 mM sodium orthovanadate – NaOVa (Na3 VO4 , Calbiochem, Cat. no. 567540), phenylarsine oxide – PAO (Sigma, Cat. no. P3075) or pervanadate – pV (obtained as described previously [ ]; using Catalase, Sigma, Cat. no. C1345 and H2 O2 Sigma, Cat. no. 31642) for 1 hour and lysed.

    Techniques: Western Blot, Expressing, shRNA, Incubation

    EGFRwt is dispensable for EGFRvIII phosphorylation ( A ) AD293 cell lines stably expressing EGFRvIII was treated with erlotinib prior to and simultaneously with NaOVa stimulation. ( B ) AD293 cell line expressing either naïve or kinase dead (KD) version of EGFRvIII on its own or together with EGFRwt have been treated with EGF or NaOVa, as indicated.

    Journal: Oncotarget

    Article Title: Cyclic trans-phosphorylation in a homodimer as the predominant mechanism of EGFRvIII action and regulation

    doi: 10.18632/oncotarget.24058

    Figure Lengend Snippet: EGFRwt is dispensable for EGFRvIII phosphorylation ( A ) AD293 cell lines stably expressing EGFRvIII was treated with erlotinib prior to and simultaneously with NaOVa stimulation. ( B ) AD293 cell line expressing either naïve or kinase dead (KD) version of EGFRvIII on its own or together with EGFRwt have been treated with EGF or NaOVa, as indicated.

    Article Snippet: Chemicals To analyse the effects of pharmacological inhibitors, cells were seeded in 6-well plates at 2.5x105 cells per well, incubated overnight to allow for adhesion and serum starved for subsequent 24 h. Serum free medium containing appropriate inhibitors was used to pre-treat cells for 30 min, unless indicated otherwise, prior to stimulation with 20 ng/mL EGF (Sigma-Aldrich, St Louis, MO, USA, Cat. no. E5036), 1 mM sodium orthovanadate – NaOVa (Na3 VO4 , Calbiochem, Cat. no. 567540), phenylarsine oxide – PAO (Sigma, Cat. no. P3075) or pervanadate – pV (obtained as described previously [ ]; using Catalase, Sigma, Cat. no. C1345 and H2 O2 Sigma, Cat. no. 31642) for 1 hour and lysed.

    Techniques: Stable Transfection, Expressing