Staining:Article Title: β 3 Integrin Promotes Long-Lasting Activation and Polarization of Vascular Endothelial Growth Factor Receptor 2 by Immobilized Ligand
Article Snippet: Adherent cells, migrating cells or VPMs underwent the following staining procedures: i) actin staining: samples were incubated for 30 minutes with TRITC-phalloidin (0.9 mg/mL in PBS, Sigma); ii) GM1 ganglioside staining: samples were incubated for 30 minutes with AlexaFluor 594- conjugated Cholera Toxin B subunit (CTB, Sigma); iii) paxillin immunostaining: samples were incubated overnight with a monoclonal anti-paxillin antibody (Transduction Laboratories, Lexington, KY) followed by a 45 minutes incubation with AlexaFluor 488-conjugated anti-mouse IgG (Molecular Probes, Life Technologies); iv) total VEGFR2 immunostaining: samples were incubated overnight with a rabbit polyclonal anti-VEGFR2 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594 and AlexaFluor 488-conjugated anti-rabbit IgG or with a goat polyclonal anti-VEGFR2 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with Alexa594-conjugated anti-goat IgG (Molecular Probes, Life Technologies); v) phospho-VEGFR2 immunostaining: samples were incubated overnight with a rabbit monoclonal anti-phosphoVEGFR2 antibody (pTyr1175, Cell Signaling Technology) followed by 1 hour incubation with AlexaFluor 488-conjugated anti-rabbit IgG; vi) β3 integrin immunostaining: samples were incubated overnight with a polyclonal anti-β3 integrin antibody (Immunological Sciences) followed by 1 hour incubation with Cy5-conjugated anti-rabbit IgG; vii) phospho-p85 immunostaining: samples were incubated overnight with a polyclonal anti-phospho-p85 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594-conjugated anti-goat IgG. viii) Caveolin 1 immunostaining: samples were incubated overnight with a mouse monoclonal anti-Caveolin 1 antibody (BD Transduction Laboratories, Franklin Lakes, New Jersey) followed by 1 hour incubation with AlexaFluor 488-conjugated anti-mouse IgG. ix) Phospho-β3 integrin immunostaining: samples were incubated overnight with a rabbit polyclonal anti-phospho-β3 integrin (pTyr759, Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594- conjugated anti-rabbit IgG.
Incubation:Article Title: β 3 Integrin Promotes Long-Lasting Activation and Polarization of Vascular Endothelial Growth Factor Receptor 2 by Immobilized Ligand
Article Snippet: Adherent cells, migrating cells or VPMs underwent the following staining procedures: i) actin staining: samples were incubated for 30 minutes with TRITC-phalloidin (0.9 mg/mL in PBS, Sigma); ii) GM1 ganglioside staining: samples were incubated for 30 minutes with AlexaFluor 594- conjugated Cholera Toxin B subunit (CTB, Sigma); iii) paxillin immunostaining: samples were incubated overnight with a monoclonal anti-paxillin antibody (Transduction Laboratories, Lexington, KY) followed by a 45 minutes incubation with AlexaFluor 488-conjugated anti-mouse IgG (Molecular Probes, Life Technologies); iv) total VEGFR2 immunostaining: samples were incubated overnight with a rabbit polyclonal anti-VEGFR2 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594 and AlexaFluor 488-conjugated anti-rabbit IgG or with a goat polyclonal anti-VEGFR2 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with Alexa594-conjugated anti-goat IgG (Molecular Probes, Life Technologies); v) phospho-VEGFR2 immunostaining: samples were incubated overnight with a rabbit monoclonal anti-phosphoVEGFR2 antibody (pTyr1175, Cell Signaling Technology) followed by 1 hour incubation with AlexaFluor 488-conjugated anti-rabbit IgG; vi) β3 integrin immunostaining: samples were incubated overnight with a polyclonal anti-β3 integrin antibody (Immunological Sciences) followed by 1 hour incubation with Cy5-conjugated anti-rabbit IgG; vii) phospho-p85 immunostaining: samples were incubated overnight with a polyclonal anti-phospho-p85 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594-conjugated anti-goat IgG. viii) Caveolin 1 immunostaining: samples were incubated overnight with a mouse monoclonal anti-Caveolin 1 antibody (BD Transduction Laboratories, Franklin Lakes, New Jersey) followed by 1 hour incubation with AlexaFluor 488-conjugated anti-mouse IgG. ix) Phospho-β3 integrin immunostaining: samples were incubated overnight with a rabbit polyclonal anti-phospho-β3 integrin (pTyr759, Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594- conjugated anti-rabbit IgG.
Immunostaining:Article Title: β 3 Integrin Promotes Long-Lasting Activation and Polarization of Vascular Endothelial Growth Factor Receptor 2 by Immobilized Ligand
Article Snippet: Adherent cells, migrating cells or VPMs underwent the following staining procedures: i) actin staining: samples were incubated for 30 minutes with TRITC-phalloidin (0.9 mg/mL in PBS, Sigma); ii) GM1 ganglioside staining: samples were incubated for 30 minutes with AlexaFluor 594- conjugated Cholera Toxin B subunit (CTB, Sigma); iii) paxillin immunostaining: samples were incubated overnight with a monoclonal anti-paxillin antibody (Transduction Laboratories, Lexington, KY) followed by a 45 minutes incubation with AlexaFluor 488-conjugated anti-mouse IgG (Molecular Probes, Life Technologies); iv) total VEGFR2 immunostaining: samples were incubated overnight with a rabbit polyclonal anti-VEGFR2 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594 and AlexaFluor 488-conjugated anti-rabbit IgG or with a goat polyclonal anti-VEGFR2 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with Alexa594-conjugated anti-goat IgG (Molecular Probes, Life Technologies); v) phospho-VEGFR2 immunostaining: samples were incubated overnight with a rabbit monoclonal anti-phosphoVEGFR2 antibody (pTyr1175, Cell Signaling Technology) followed by 1 hour incubation with AlexaFluor 488-conjugated anti-rabbit IgG; vi) β3 integrin immunostaining: samples were incubated overnight with a polyclonal anti-β3 integrin antibody (Immunological Sciences) followed by 1 hour incubation with Cy5-conjugated anti-rabbit IgG; vii) phospho-p85 immunostaining: samples were incubated overnight with a polyclonal anti-phospho-p85 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594-conjugated anti-goat IgG. viii) Caveolin 1 immunostaining: samples were incubated overnight with a mouse monoclonal anti-Caveolin 1 antibody (BD Transduction Laboratories, Franklin Lakes, New Jersey) followed by 1 hour incubation with AlexaFluor 488-conjugated anti-mouse IgG. ix) Phospho-β3 integrin immunostaining: samples were incubated overnight with a rabbit polyclonal anti-phospho-β3 integrin (pTyr759, Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594- conjugated anti-rabbit IgG.
Transduction:Article Title: β 3 Integrin Promotes Long-Lasting Activation and Polarization of Vascular Endothelial Growth Factor Receptor 2 by Immobilized Ligand
Article Snippet: Adherent cells, migrating cells or VPMs underwent the following staining procedures: i) actin staining: samples were incubated for 30 minutes with TRITC-phalloidin (0.9 mg/mL in PBS, Sigma); ii) GM1 ganglioside staining: samples were incubated for 30 minutes with AlexaFluor 594- conjugated Cholera Toxin B subunit (CTB, Sigma); iii) paxillin immunostaining: samples were incubated overnight with a monoclonal anti-paxillin antibody (Transduction Laboratories, Lexington, KY) followed by a 45 minutes incubation with AlexaFluor 488-conjugated anti-mouse IgG (Molecular Probes, Life Technologies); iv) total VEGFR2 immunostaining: samples were incubated overnight with a rabbit polyclonal anti-VEGFR2 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594 and AlexaFluor 488-conjugated anti-rabbit IgG or with a goat polyclonal anti-VEGFR2 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with Alexa594-conjugated anti-goat IgG (Molecular Probes, Life Technologies); v) phospho-VEGFR2 immunostaining: samples were incubated overnight with a rabbit monoclonal anti-phosphoVEGFR2 antibody (pTyr1175, Cell Signaling Technology) followed by 1 hour incubation with AlexaFluor 488-conjugated anti-rabbit IgG; vi) β3 integrin immunostaining: samples were incubated overnight with a polyclonal anti-β3 integrin antibody (Immunological Sciences) followed by 1 hour incubation with Cy5-conjugated anti-rabbit IgG; vii) phospho-p85 immunostaining: samples were incubated overnight with a polyclonal anti-phospho-p85 antibody (Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594-conjugated anti-goat IgG. viii) Caveolin 1 immunostaining: samples were incubated overnight with a mouse monoclonal anti-Caveolin 1 antibody (BD Transduction Laboratories, Franklin Lakes, New Jersey) followed by 1 hour incubation with AlexaFluor 488-conjugated anti-mouse IgG. ix) Phospho-β3 integrin immunostaining: samples were incubated overnight with a rabbit polyclonal anti-phospho-β3 integrin (pTyr759, Santa Cruz Biotechnology) followed by 1 hour incubation with AlexaFluor 594- conjugated anti-rabbit IgG.
|
Santa Cruz Biotechnology is a verified supplier 