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phospho-erbb4 (tyr 1188  (Orbigen Inc)

 
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    Orbigen Inc phospho-erbb4 (tyr 1188
    Activation of the <t>ErbB4</t> RTK in ET spheroids. A, the R&D Systems Human Phospho-RTK Antibody Proteome Profiler Array system was used to screen for activation of specific tyrosine kinases in ET spheroids. Lysates from TC32 and TC71 monolayer and spheroid cultures were incubated with membranes arrayed with antibodies from 42 different tyrosine kinases (http://www.rndsystems.com/). Membranes were then washed and incubated with anti–phosphotyrosine-HRP antibodies followed by enhanced chemiluminescence detection to identify activated tyrosine kinases. P-Tyr, phospho-tyrosine control spots. B, top, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation (IP) with 4G10 anti-phosphotyrosine antibodies and then immunoblotted (IB) with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. n.s., nonspecific band. Left, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation with antibodies to total ErbB4 followed by immunoblotting with anti-phosphotyrosine antibodies. Right, the blot was then reprobed with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. C, TC32 and TC71 cells were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188). Detection of β-actin was used as a loading control. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. D, ErbB4 and E-cadherin expression was analyzed by immunohistochemistry from sections from primary ET. Sections were subjected to standard H&E staining, as well as immunohistochemistry with anti-ErbB4 (ErbB4), anti–E-cadherin, and control IgG antibodies. Magnification, ×400.
    Phospho Erbb4 (Tyr 1188, supplied by Orbigen Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho-erbb4 (tyr 1188/product/Orbigen Inc
    Average 90 stars, based on 1 article reviews
    phospho-erbb4 (tyr 1188 - by Bioz Stars, 2026-03
    90/100 stars

    Images

    1) Product Images from "E-Cadherin Cell-Cell Adhesion in Ewing Tumor Cells Mediates Suppression of Anoikis through Activation of the ErbB4 Tyrosine Kinase"

    Article Title: E-Cadherin Cell-Cell Adhesion in Ewing Tumor Cells Mediates Suppression of Anoikis through Activation of the ErbB4 Tyrosine Kinase

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-06-3259

    Activation of the ErbB4 RTK in ET spheroids. A, the R&D Systems Human Phospho-RTK Antibody Proteome Profiler Array system was used to screen for activation of specific tyrosine kinases in ET spheroids. Lysates from TC32 and TC71 monolayer and spheroid cultures were incubated with membranes arrayed with antibodies from 42 different tyrosine kinases (http://www.rndsystems.com/). Membranes were then washed and incubated with anti–phosphotyrosine-HRP antibodies followed by enhanced chemiluminescence detection to identify activated tyrosine kinases. P-Tyr, phospho-tyrosine control spots. B, top, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation (IP) with 4G10 anti-phosphotyrosine antibodies and then immunoblotted (IB) with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. n.s., nonspecific band. Left, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation with antibodies to total ErbB4 followed by immunoblotting with anti-phosphotyrosine antibodies. Right, the blot was then reprobed with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. C, TC32 and TC71 cells were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188). Detection of β-actin was used as a loading control. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. D, ErbB4 and E-cadherin expression was analyzed by immunohistochemistry from sections from primary ET. Sections were subjected to standard H&E staining, as well as immunohistochemistry with anti-ErbB4 (ErbB4), anti–E-cadherin, and control IgG antibodies. Magnification, ×400.
    Figure Legend Snippet: Activation of the ErbB4 RTK in ET spheroids. A, the R&D Systems Human Phospho-RTK Antibody Proteome Profiler Array system was used to screen for activation of specific tyrosine kinases in ET spheroids. Lysates from TC32 and TC71 monolayer and spheroid cultures were incubated with membranes arrayed with antibodies from 42 different tyrosine kinases (http://www.rndsystems.com/). Membranes were then washed and incubated with anti–phosphotyrosine-HRP antibodies followed by enhanced chemiluminescence detection to identify activated tyrosine kinases. P-Tyr, phospho-tyrosine control spots. B, top, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation (IP) with 4G10 anti-phosphotyrosine antibodies and then immunoblotted (IB) with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. n.s., nonspecific band. Left, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation with antibodies to total ErbB4 followed by immunoblotting with anti-phosphotyrosine antibodies. Right, the blot was then reprobed with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. C, TC32 and TC71 cells were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188). Detection of β-actin was used as a loading control. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. D, ErbB4 and E-cadherin expression was analyzed by immunohistochemistry from sections from primary ET. Sections were subjected to standard H&E staining, as well as immunohistochemistry with anti-ErbB4 (ErbB4), anti–E-cadherin, and control IgG antibodies. Magnification, ×400.

    Techniques Used: Activation Assay, Incubation, Immunoprecipitation, Western Blot, Expressing, Immunohistochemistry, Staining

    ErbB4 RNA interference blocks Akt activation and restores chemosensitivity of ET spheroids. A, TC32 and TC71 cell lines that were either nontransduced or stably expressing DN-Ecad or its vector alone or Ecad or its vector alone were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188; p-ErbB4) or total ErbB4. B, ErbB4 knockdown with RNA interference blocks Akt activation. TC32 spheroids grown in media containing 10% serum were treated with two ErbB4-specific siRNA pools (siRNA1 or siRNA2), as described in Materials and Methods, or a scrambled siRNA control. Lysates were then subjected to immunoblotting with antibodies to total ErbB4, phospho-Akt (pAKT), phospho-MEK (pMEK), or β-actin as a loading control. C, phase-contrast photomicrographs of TC32 spheroids after pretreatment of cells with an ErbB4-specific siRNA (siRNA1) or a scrambled siRNA control. Magnification, ×40.D, reduction of ErbB4 by RNA interference restores chemosensitivity of ET spheroids. TC32 spheroids grown in media containing 10% serum were treated with ErbB4-specific siRNAs or a scrambled control as above. Cells were then treated with vehicle control, 50 µmol/L etoposide, or 10 µmol/L doxorubicin for 24 h after siRNA transfection. Caspase-3 activity was then assayed as above with normalization to a value of 1.0 arbitrary unit for nontreated cells. Statistical analysis of data from at least three separate experiments was done using Student's t test. Horizontal lines, P values comparing scrambled siRNA control with ErbB4 siRNA1 cells in the presence of etoposide (P < 0.004) or doxorubicin (P < 0.009), or comparing scrambled siRNA control with ErbB4 siRNA2 in the presence of etoposide (P < 0.0006) or doxorubicin (P < 0.003).
    Figure Legend Snippet: ErbB4 RNA interference blocks Akt activation and restores chemosensitivity of ET spheroids. A, TC32 and TC71 cell lines that were either nontransduced or stably expressing DN-Ecad or its vector alone or Ecad or its vector alone were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188; p-ErbB4) or total ErbB4. B, ErbB4 knockdown with RNA interference blocks Akt activation. TC32 spheroids grown in media containing 10% serum were treated with two ErbB4-specific siRNA pools (siRNA1 or siRNA2), as described in Materials and Methods, or a scrambled siRNA control. Lysates were then subjected to immunoblotting with antibodies to total ErbB4, phospho-Akt (pAKT), phospho-MEK (pMEK), or β-actin as a loading control. C, phase-contrast photomicrographs of TC32 spheroids after pretreatment of cells with an ErbB4-specific siRNA (siRNA1) or a scrambled siRNA control. Magnification, ×40.D, reduction of ErbB4 by RNA interference restores chemosensitivity of ET spheroids. TC32 spheroids grown in media containing 10% serum were treated with ErbB4-specific siRNAs or a scrambled control as above. Cells were then treated with vehicle control, 50 µmol/L etoposide, or 10 µmol/L doxorubicin for 24 h after siRNA transfection. Caspase-3 activity was then assayed as above with normalization to a value of 1.0 arbitrary unit for nontreated cells. Statistical analysis of data from at least three separate experiments was done using Student's t test. Horizontal lines, P values comparing scrambled siRNA control with ErbB4 siRNA1 cells in the presence of etoposide (P < 0.004) or doxorubicin (P < 0.009), or comparing scrambled siRNA control with ErbB4 siRNA2 in the presence of etoposide (P < 0.0006) or doxorubicin (P < 0.003).

    Techniques Used: Activation Assay, Stable Transfection, Expressing, Plasmid Preparation, Western Blot, Transfection, Activity Assay



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    phospho-erbb4 (tyr 1188  (Orbigen Inc)
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    Orbigen Inc phospho-erbb4 (tyr 1188
    Activation of the <t>ErbB4</t> RTK in ET spheroids. A, the R&D Systems Human Phospho-RTK Antibody Proteome Profiler Array system was used to screen for activation of specific tyrosine kinases in ET spheroids. Lysates from TC32 and TC71 monolayer and spheroid cultures were incubated with membranes arrayed with antibodies from 42 different tyrosine kinases (http://www.rndsystems.com/). Membranes were then washed and incubated with anti–phosphotyrosine-HRP antibodies followed by enhanced chemiluminescence detection to identify activated tyrosine kinases. P-Tyr, phospho-tyrosine control spots. B, top, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation (IP) with 4G10 anti-phosphotyrosine antibodies and then immunoblotted (IB) with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. n.s., nonspecific band. Left, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation with antibodies to total ErbB4 followed by immunoblotting with anti-phosphotyrosine antibodies. Right, the blot was then reprobed with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. C, TC32 and TC71 cells were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188). Detection of β-actin was used as a loading control. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. D, ErbB4 and E-cadherin expression was analyzed by immunohistochemistry from sections from primary ET. Sections were subjected to standard H&E staining, as well as immunohistochemistry with anti-ErbB4 (ErbB4), anti–E-cadherin, and control IgG antibodies. Magnification, ×400.
    Phospho Erbb4 (Tyr 1188, supplied by Orbigen Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho-erbb4 (tyr 1188/product/Orbigen Inc
    Average 90 stars, based on 1 article reviews
    phospho-erbb4 (tyr 1188 - by Bioz Stars, 2026-03
    90/100 stars
    		  Buy from Supplier

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    Activation of the ErbB4 RTK in ET spheroids. A, the R&D Systems Human Phospho-RTK Antibody Proteome Profiler Array system was used to screen for activation of specific tyrosine kinases in ET spheroids. Lysates from TC32 and TC71 monolayer and spheroid cultures were incubated with membranes arrayed with antibodies from 42 different tyrosine kinases (http://www.rndsystems.com/). Membranes were then washed and incubated with anti–phosphotyrosine-HRP antibodies followed by enhanced chemiluminescence detection to identify activated tyrosine kinases. P-Tyr, phospho-tyrosine control spots. B, top, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation (IP) with 4G10 anti-phosphotyrosine antibodies and then immunoblotted (IB) with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. n.s., nonspecific band. Left, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation with antibodies to total ErbB4 followed by immunoblotting with anti-phosphotyrosine antibodies. Right, the blot was then reprobed with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. C, TC32 and TC71 cells were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188). Detection of β-actin was used as a loading control. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. D, ErbB4 and E-cadherin expression was analyzed by immunohistochemistry from sections from primary ET. Sections were subjected to standard H&E staining, as well as immunohistochemistry with anti-ErbB4 (ErbB4), anti–E-cadherin, and control IgG antibodies. Magnification, ×400.

    Journal: Cancer research

    Article Title: E-Cadherin Cell-Cell Adhesion in Ewing Tumor Cells Mediates Suppression of Anoikis through Activation of the ErbB4 Tyrosine Kinase

    doi: 10.1158/0008-5472.CAN-06-3259

    Figure Lengend Snippet: Activation of the ErbB4 RTK in ET spheroids. A, the R&D Systems Human Phospho-RTK Antibody Proteome Profiler Array system was used to screen for activation of specific tyrosine kinases in ET spheroids. Lysates from TC32 and TC71 monolayer and spheroid cultures were incubated with membranes arrayed with antibodies from 42 different tyrosine kinases (http://www.rndsystems.com/). Membranes were then washed and incubated with anti–phosphotyrosine-HRP antibodies followed by enhanced chemiluminescence detection to identify activated tyrosine kinases. P-Tyr, phospho-tyrosine control spots. B, top, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation (IP) with 4G10 anti-phosphotyrosine antibodies and then immunoblotted (IB) with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. n.s., nonspecific band. Left, lysates from TC32 and TC71 monolayer and spheroid cultures were subjected to immunoprecipitation with antibodies to total ErbB4 followed by immunoblotting with anti-phosphotyrosine antibodies. Right, the blot was then reprobed with an anti-ErbB4 antibody. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. C, TC32 and TC71 cells were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188). Detection of β-actin was used as a loading control. The positions of 180- and 80-kDa tyrosine phosphorylated species are indicated. D, ErbB4 and E-cadherin expression was analyzed by immunohistochemistry from sections from primary ET. Sections were subjected to standard H&E staining, as well as immunohistochemistry with anti-ErbB4 (ErbB4), anti–E-cadherin, and control IgG antibodies. Magnification, ×400.

    Article Snippet: Standard Western blot analysis was done with antibodies to poly(ADP-ribose) polymerase, phospho-Akt Ser 473 , total Akt, phospho–mitogen-activated protein kinase/ERK kinase (MEK) 1/2 Ser 217/221 (Cell Signaling, Beverly, MA); β-actin, ErbB4, ErbB2 (Santa Cruz Biotechnology, Santa Cruz, CA); phospho-ErbB4 (Tyr 1188 ), phospho-ErbB2 (Tyr 1112 ) (Orbigen, San Diego, CA); E-cadherin, Rac1 (BD Transduction Laboratories, San Diego, CA); and phosphotyrosine (4G10 from Upstate, Lake Placid, NY).

    Techniques: Activation Assay, Incubation, Immunoprecipitation, Western Blot, Expressing, Immunohistochemistry, Staining

    ErbB4 RNA interference blocks Akt activation and restores chemosensitivity of ET spheroids. A, TC32 and TC71 cell lines that were either nontransduced or stably expressing DN-Ecad or its vector alone or Ecad or its vector alone were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188; p-ErbB4) or total ErbB4. B, ErbB4 knockdown with RNA interference blocks Akt activation. TC32 spheroids grown in media containing 10% serum were treated with two ErbB4-specific siRNA pools (siRNA1 or siRNA2), as described in Materials and Methods, or a scrambled siRNA control. Lysates were then subjected to immunoblotting with antibodies to total ErbB4, phospho-Akt (pAKT), phospho-MEK (pMEK), or β-actin as a loading control. C, phase-contrast photomicrographs of TC32 spheroids after pretreatment of cells with an ErbB4-specific siRNA (siRNA1) or a scrambled siRNA control. Magnification, ×40.D, reduction of ErbB4 by RNA interference restores chemosensitivity of ET spheroids. TC32 spheroids grown in media containing 10% serum were treated with ErbB4-specific siRNAs or a scrambled control as above. Cells were then treated with vehicle control, 50 µmol/L etoposide, or 10 µmol/L doxorubicin for 24 h after siRNA transfection. Caspase-3 activity was then assayed as above with normalization to a value of 1.0 arbitrary unit for nontreated cells. Statistical analysis of data from at least three separate experiments was done using Student's t test. Horizontal lines, P values comparing scrambled siRNA control with ErbB4 siRNA1 cells in the presence of etoposide (P < 0.004) or doxorubicin (P < 0.009), or comparing scrambled siRNA control with ErbB4 siRNA2 in the presence of etoposide (P < 0.0006) or doxorubicin (P < 0.003).

    Journal: Cancer research

    Article Title: E-Cadherin Cell-Cell Adhesion in Ewing Tumor Cells Mediates Suppression of Anoikis through Activation of the ErbB4 Tyrosine Kinase

    doi: 10.1158/0008-5472.CAN-06-3259

    Figure Lengend Snippet: ErbB4 RNA interference blocks Akt activation and restores chemosensitivity of ET spheroids. A, TC32 and TC71 cell lines that were either nontransduced or stably expressing DN-Ecad or its vector alone or Ecad or its vector alone were grown as monolayers or spheroids as indicated in 10% serum–containing media for 48 h. Cells were then lysed and subjected to immunoblotting with antibodies to activated ErbB4 (phospho-Tyr1188; p-ErbB4) or total ErbB4. B, ErbB4 knockdown with RNA interference blocks Akt activation. TC32 spheroids grown in media containing 10% serum were treated with two ErbB4-specific siRNA pools (siRNA1 or siRNA2), as described in Materials and Methods, or a scrambled siRNA control. Lysates were then subjected to immunoblotting with antibodies to total ErbB4, phospho-Akt (pAKT), phospho-MEK (pMEK), or β-actin as a loading control. C, phase-contrast photomicrographs of TC32 spheroids after pretreatment of cells with an ErbB4-specific siRNA (siRNA1) or a scrambled siRNA control. Magnification, ×40.D, reduction of ErbB4 by RNA interference restores chemosensitivity of ET spheroids. TC32 spheroids grown in media containing 10% serum were treated with ErbB4-specific siRNAs or a scrambled control as above. Cells were then treated with vehicle control, 50 µmol/L etoposide, or 10 µmol/L doxorubicin for 24 h after siRNA transfection. Caspase-3 activity was then assayed as above with normalization to a value of 1.0 arbitrary unit for nontreated cells. Statistical analysis of data from at least three separate experiments was done using Student's t test. Horizontal lines, P values comparing scrambled siRNA control with ErbB4 siRNA1 cells in the presence of etoposide (P < 0.004) or doxorubicin (P < 0.009), or comparing scrambled siRNA control with ErbB4 siRNA2 in the presence of etoposide (P < 0.0006) or doxorubicin (P < 0.003).

    Article Snippet: Standard Western blot analysis was done with antibodies to poly(ADP-ribose) polymerase, phospho-Akt Ser 473 , total Akt, phospho–mitogen-activated protein kinase/ERK kinase (MEK) 1/2 Ser 217/221 (Cell Signaling, Beverly, MA); β-actin, ErbB4, ErbB2 (Santa Cruz Biotechnology, Santa Cruz, CA); phospho-ErbB4 (Tyr 1188 ), phospho-ErbB2 (Tyr 1112 ) (Orbigen, San Diego, CA); E-cadherin, Rac1 (BD Transduction Laboratories, San Diego, CA); and phosphotyrosine (4G10 from Upstate, Lake Placid, NY).

    Techniques: Activation Assay, Stable Transfection, Expressing, Plasmid Preparation, Western Blot, Transfection, Activity Assay