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Clinicopathological parameters for the clinical tumour set
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Clinicopathological parameters for the clinical tumour set

Journal: Breast cancer research : BCR

Article Title: Heregulin β1 drives gefitinib-resistant growth and invasion in tamoxifen-resistant MCF-7 breast cancer cells

doi: 10.1186/bcr1754

Figure Lengend Snippet: Clinicopathological parameters for the clinical tumour set

Article Snippet: The antibodies used were total EGFR (SC-03) erbB2 (SC-284), erbB3 (SC-285) and erbB4 (SC-283) (Insight Biotechnology Ltd), anti-phospho-erbB2 (pY1248, 2247), anti-phospho-EGFR (pY1068, 2234), total AKT (9272), phospho-AKT (pS473, 9271), total ERK1/2 (9102) and phospho-ERK1/2 (pT202/pY204, 9101) (New England Biolabs, Hitchin, Hertfordshire, UK), and β-actin (AC-15) (Sigma).

Techniques:

Effects of gefitinib on erbB receptor dimerization patterns and activity of associated downstream signalling elements. (a) Western blot (WB) analysis of phosphorylated epidermal growth factor receptor (p-EGFR), phosphorylated erbB2, phosphorylated tyrosine (Tyr) and total erbB3 protein expression following immunoprecipitation with total erbB3 antibody in tamoxifen-resistant MCF-7 (Tam-R) cells prior to and following incubation of Tam-R cells with gefitinib (1 μM) for 1 hour. (b) WB analysis of total and phosphorylated EGFR, erbB2, AKT and ERK1/2 protein expression in Tam-R cells prior to and following incubation with gefitinib (1 μM) for 1 hour. (c) WB analysis of total and phosphorylated EGFR, erbB2 and erbB3 protein expression following immunoprecipitation with total EGFR antibody in Tam-R cells prior to and following incubation with gefitinib (1 μM) for 1 hour. Tamoxifen (100 nM) was present in all studies. Data are representative of at least three separate experiments.

Journal: Breast cancer research : BCR

Article Title: Heregulin β1 drives gefitinib-resistant growth and invasion in tamoxifen-resistant MCF-7 breast cancer cells

doi: 10.1186/bcr1754

Figure Lengend Snippet: Effects of gefitinib on erbB receptor dimerization patterns and activity of associated downstream signalling elements. (a) Western blot (WB) analysis of phosphorylated epidermal growth factor receptor (p-EGFR), phosphorylated erbB2, phosphorylated tyrosine (Tyr) and total erbB3 protein expression following immunoprecipitation with total erbB3 antibody in tamoxifen-resistant MCF-7 (Tam-R) cells prior to and following incubation of Tam-R cells with gefitinib (1 μM) for 1 hour. (b) WB analysis of total and phosphorylated EGFR, erbB2, AKT and ERK1/2 protein expression in Tam-R cells prior to and following incubation with gefitinib (1 μM) for 1 hour. (c) WB analysis of total and phosphorylated EGFR, erbB2 and erbB3 protein expression following immunoprecipitation with total EGFR antibody in Tam-R cells prior to and following incubation with gefitinib (1 μM) for 1 hour. Tamoxifen (100 nM) was present in all studies. Data are representative of at least three separate experiments.

Article Snippet: The antibodies used were total EGFR (SC-03) erbB2 (SC-284), erbB3 (SC-285) and erbB4 (SC-283) (Insight Biotechnology Ltd), anti-phospho-erbB2 (pY1248, 2247), anti-phospho-EGFR (pY1068, 2234), total AKT (9272), phospho-AKT (pS473, 9271), total ERK1/2 (9102) and phospho-ERK1/2 (pT202/pY204, 9101) (New England Biolabs, Hitchin, Hertfordshire, UK), and β-actin (AC-15) (Sigma).

Techniques: Activity Assay, Western Blot, Expressing, Immunoprecipitation, Incubation

Effects of HRGβ1 and gefitinib on erbB receptor dimerization patterns and associated downstream signalling activity. (a) Western blot (WB) analysis of phosphorylated epidermal growth factor receptor (EGFR), phosphorylated erbB2, phosphorylated tyrosine (Tyr) and total erbB3 protein expression following immunoprecipitation with total erbB3 antibody in tamoxifen-resistant MCF-7 (Tam-R) cells prior to and following treatment with either gefitinib (1 μM) or vehicle control for 1 hour followed by HRGβ1 (10 ng/ml) for 5 minutes. (b) WB analysis of total and phosphorylated EGFR, erbB2, AKT and ERK1/2 protein expression in Tam-R cells prior to and following incubation with either gefitinib (1 μM) or vehicle control for 1 hour followed by either HRGβ1 (10 ng/ml) or vehicle control for 5 minutes. Tamoxifen was also present in all studies. Data are representative of at least three separate experiments.

Journal: Breast cancer research : BCR

Article Title: Heregulin β1 drives gefitinib-resistant growth and invasion in tamoxifen-resistant MCF-7 breast cancer cells

doi: 10.1186/bcr1754

Figure Lengend Snippet: Effects of HRGβ1 and gefitinib on erbB receptor dimerization patterns and associated downstream signalling activity. (a) Western blot (WB) analysis of phosphorylated epidermal growth factor receptor (EGFR), phosphorylated erbB2, phosphorylated tyrosine (Tyr) and total erbB3 protein expression following immunoprecipitation with total erbB3 antibody in tamoxifen-resistant MCF-7 (Tam-R) cells prior to and following treatment with either gefitinib (1 μM) or vehicle control for 1 hour followed by HRGβ1 (10 ng/ml) for 5 minutes. (b) WB analysis of total and phosphorylated EGFR, erbB2, AKT and ERK1/2 protein expression in Tam-R cells prior to and following incubation with either gefitinib (1 μM) or vehicle control for 1 hour followed by either HRGβ1 (10 ng/ml) or vehicle control for 5 minutes. Tamoxifen was also present in all studies. Data are representative of at least three separate experiments.

Article Snippet: The antibodies used were total EGFR (SC-03) erbB2 (SC-284), erbB3 (SC-285) and erbB4 (SC-283) (Insight Biotechnology Ltd), anti-phospho-erbB2 (pY1248, 2247), anti-phospho-EGFR (pY1068, 2234), total AKT (9272), phospho-AKT (pS473, 9271), total ERK1/2 (9102) and phospho-ERK1/2 (pT202/pY204, 9101) (New England Biolabs, Hitchin, Hertfordshire, UK), and β-actin (AC-15) (Sigma).

Techniques: Activity Assay, Western Blot, Expressing, Immunoprecipitation, Incubation

Effects of combining gefitinib with trastuzumab or LY294002 on HRGβ1-driven signalling in tamoxifen-resistant MCF-7 cells. Western analysis of total and phosphorylated epidermal growth factor receptor (EGFR), erbB2, AKT and ERK1/2 protein expression in tamoxifen-resistant MCF-7 (Tam-R) cells prior to and following incubation with either (a) trastuzumab (100 nM) or vehicle control for 7 days, (b) LY294002 (10 μM) or vehicle control for 1 hour, (c) gefitinib (1 μM), gefitinib in combination with trastuzumab or vehicle control for 7 days, and (d) gefitinib, gefitinib in combination with LY294002 or vehicle control for 1 hour, all followed by either HRGβ1 (10 ng/ml) or vehicle control for 5 minutes. Tamoxifen was also present in all studies. Data are representative of at least three separate experiments.

Journal: Breast cancer research : BCR

Article Title: Heregulin β1 drives gefitinib-resistant growth and invasion in tamoxifen-resistant MCF-7 breast cancer cells

doi: 10.1186/bcr1754

Figure Lengend Snippet: Effects of combining gefitinib with trastuzumab or LY294002 on HRGβ1-driven signalling in tamoxifen-resistant MCF-7 cells. Western analysis of total and phosphorylated epidermal growth factor receptor (EGFR), erbB2, AKT and ERK1/2 protein expression in tamoxifen-resistant MCF-7 (Tam-R) cells prior to and following incubation with either (a) trastuzumab (100 nM) or vehicle control for 7 days, (b) LY294002 (10 μM) or vehicle control for 1 hour, (c) gefitinib (1 μM), gefitinib in combination with trastuzumab or vehicle control for 7 days, and (d) gefitinib, gefitinib in combination with LY294002 or vehicle control for 1 hour, all followed by either HRGβ1 (10 ng/ml) or vehicle control for 5 minutes. Tamoxifen was also present in all studies. Data are representative of at least three separate experiments.

Article Snippet: The antibodies used were total EGFR (SC-03) erbB2 (SC-284), erbB3 (SC-285) and erbB4 (SC-283) (Insight Biotechnology Ltd), anti-phospho-erbB2 (pY1248, 2247), anti-phospho-EGFR (pY1068, 2234), total AKT (9272), phospho-AKT (pS473, 9271), total ERK1/2 (9102) and phospho-ERK1/2 (pT202/pY204, 9101) (New England Biolabs, Hitchin, Hertfordshire, UK), and β-actin (AC-15) (Sigma).

Techniques: Western Blot, Expressing, Incubation

Immunohistochemical staining for HRGβ1 in primary breast cancer specimens. (a) Examples of high and low HRGβ1 expression. Scale bars = 20 μm. (b) Box-plots illustrating cytoplasmic HRGβ1 immunohistochemistry assessed by H-score analysis in membrane (mem) epidermal growth factor receptor (EGFR)-negative, erbB2-negative and erbB3-negative primary breast cancer versus membrane EGFR-positive, erbB2-positive and erbB3-positive primary breast cancer. A significant positive correlation between cytoplasmic HRGβ1 expression and membrane erbB receptor positivity was only seen with EGFR (Mann–Whitney U test, P = 0.04). (c) Box-plots illustrating cytoplasmic HRGβ1 immunohistochemistry assessed by H-score analysis in membrane (mem) phosphorylated erbB2-negative versus membrane phosphorylated erbB2-positive primary breast cancer and in nuclear (nuc) phosphorylated ERK1/2-negative versus nuclear phosphorylated ERK1/2-positive primary breast cancer (Mann–Whitney U test, P = 0.006 and P = 0.017, respectively), and scatter plot illustrating the significant positive correlation between expression levels of nuclear phosphorylated AKT and cytoplasmic HRGβ1 in the same samples (Spearman rank test, P = 0.044).

Journal: Breast cancer research : BCR

Article Title: Heregulin β1 drives gefitinib-resistant growth and invasion in tamoxifen-resistant MCF-7 breast cancer cells

doi: 10.1186/bcr1754

Figure Lengend Snippet: Immunohistochemical staining for HRGβ1 in primary breast cancer specimens. (a) Examples of high and low HRGβ1 expression. Scale bars = 20 μm. (b) Box-plots illustrating cytoplasmic HRGβ1 immunohistochemistry assessed by H-score analysis in membrane (mem) epidermal growth factor receptor (EGFR)-negative, erbB2-negative and erbB3-negative primary breast cancer versus membrane EGFR-positive, erbB2-positive and erbB3-positive primary breast cancer. A significant positive correlation between cytoplasmic HRGβ1 expression and membrane erbB receptor positivity was only seen with EGFR (Mann–Whitney U test, P = 0.04). (c) Box-plots illustrating cytoplasmic HRGβ1 immunohistochemistry assessed by H-score analysis in membrane (mem) phosphorylated erbB2-negative versus membrane phosphorylated erbB2-positive primary breast cancer and in nuclear (nuc) phosphorylated ERK1/2-negative versus nuclear phosphorylated ERK1/2-positive primary breast cancer (Mann–Whitney U test, P = 0.006 and P = 0.017, respectively), and scatter plot illustrating the significant positive correlation between expression levels of nuclear phosphorylated AKT and cytoplasmic HRGβ1 in the same samples (Spearman rank test, P = 0.044).

Article Snippet: The antibodies used were total EGFR (SC-03) erbB2 (SC-284), erbB3 (SC-285) and erbB4 (SC-283) (Insight Biotechnology Ltd), anti-phospho-erbB2 (pY1248, 2247), anti-phospho-EGFR (pY1068, 2234), total AKT (9272), phospho-AKT (pS473, 9271), total ERK1/2 (9102) and phospho-ERK1/2 (pT202/pY204, 9101) (New England Biolabs, Hitchin, Hertfordshire, UK), and β-actin (AC-15) (Sigma).

Techniques: Immunohistochemical staining, Staining, Expressing, Immunohistochemistry, MANN-WHITNEY