ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ser473
    BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT <t>(Ser473),</t> total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.
    Ser473, 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
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    Average 94 stars, based on 1 article reviews
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    1) Product Images from "BCAP31 drives TNBC development by modulating ligand-independent EGFR trafficking and spontaneous EGFR phosphorylation"

    Article Title: BCAP31 drives TNBC development by modulating ligand-independent EGFR trafficking and spontaneous EGFR phosphorylation

    Journal: Theranostics

    doi: 10.7150/thno.35383

    BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT (Ser473), total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.
    Figure Legend Snippet: BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT (Ser473), total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.

    Techniques Used: Activation Assay, Mass Spectrometry, Purification, SDS Page, Silver Staining, Co-Immunoprecipitation Assay, Western Blot, Stable Transfection, shRNA, Expressing, Infection, Enzyme-linked Immunosorbent Assay

    ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ser473
    BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT <t>(Ser473),</t> total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.
    Ser473, 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
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    Average 94 stars, based on 1 article reviews
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    ser473 - by Bioz Stars, 2023-01
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    Images

    1) Product Images from "BCAP31 drives TNBC development by modulating ligand-independent EGFR trafficking and spontaneous EGFR phosphorylation"

    Article Title: BCAP31 drives TNBC development by modulating ligand-independent EGFR trafficking and spontaneous EGFR phosphorylation

    Journal: Theranostics

    doi: 10.7150/thno.35383

    BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT (Ser473), total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.
    Figure Legend Snippet: BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT (Ser473), total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.

    Techniques Used: Activation Assay, Mass Spectrometry, Purification, SDS Page, Silver Staining, Co-Immunoprecipitation Assay, Western Blot, Stable Transfection, shRNA, Expressing, Infection, Enzyme-linked Immunosorbent Assay

    p akt1 2 3 ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p akt1 2 3 ser473
    P Akt1 2 3 Ser473, 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
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    pakt s473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt s473
    ( A ) Phalloidin detection shows cells with polarized F-actin (white arrow) following treatment with recombinant Periostin compared to vehicle control. Scale bars represent 10 μm. ( B ) Graphic represents percentage of cells with stress fiber formation (polarized F-actin) after periostin or vehicle stimuli. Results were determined by measuring fields using independent triplicates (**p<0.01) ( C ) Activation of PI3K and mTOR signaling is triggered by Periostin treatment in a dose-dependent manner, as detected by phosphorylated AKT at Threonine 308 <t>(pAKT</t> Thr308 ) and <t>Serine</t> <t>473</t> (pAKT Ser473 ) and phosphorylated S6 (pS6). Note that 50 ng/ml of Periostin is the optimal concentration for PI3K activation. GAPDH was used as a loading control.
    Pakt S473, 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
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    1) Product Images from "Periostin Responds to Mechanical Stress and Tension by Activating the MTOR Signaling Pathway"

    Article Title: Periostin Responds to Mechanical Stress and Tension by Activating the MTOR Signaling Pathway

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0083580

    ( A ) Phalloidin detection shows cells with polarized F-actin (white arrow) following treatment with recombinant Periostin compared to vehicle control. Scale bars represent 10 μm. ( B ) Graphic represents percentage of cells with stress fiber formation (polarized F-actin) after periostin or vehicle stimuli. Results were determined by measuring fields using independent triplicates (**p<0.01) ( C ) Activation of PI3K and mTOR signaling is triggered by Periostin treatment in a dose-dependent manner, as detected by phosphorylated AKT at Threonine 308 (pAKT Thr308 ) and Serine 473 (pAKT Ser473 ) and phosphorylated S6 (pS6). Note that 50 ng/ml of Periostin is the optimal concentration for PI3K activation. GAPDH was used as a loading control.
    Figure Legend Snippet: ( A ) Phalloidin detection shows cells with polarized F-actin (white arrow) following treatment with recombinant Periostin compared to vehicle control. Scale bars represent 10 μm. ( B ) Graphic represents percentage of cells with stress fiber formation (polarized F-actin) after periostin or vehicle stimuli. Results were determined by measuring fields using independent triplicates (**p<0.01) ( C ) Activation of PI3K and mTOR signaling is triggered by Periostin treatment in a dose-dependent manner, as detected by phosphorylated AKT at Threonine 308 (pAKT Thr308 ) and Serine 473 (pAKT Ser473 ) and phosphorylated S6 (pS6). Note that 50 ng/ml of Periostin is the optimal concentration for PI3K activation. GAPDH was used as a loading control.

    Techniques Used: Recombinant, Activation Assay, Concentration Assay

    p akt1 ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p akt1 ser473
    KLK5 rescued the abrogated GBC progression in IGF2BP3-depleted cells. (A–E) IGF2BP3-depleted GBC cells were co-transfected with PLVX-KLK5 or control plasmid. Cell growth ability was assessed by CCK8 assay (A) , colony formation assay (B) and EdU assay ( C , 100X, scale bar: 100μm). Statistical significance was analyzed based on the number of EdU stained cells (right). (D) Transwell assays were conducted to compare the migration ability in each treated group of GBC cells (100X, scale bar: 100μm). (E) Western blotting assays showed the expression of IGF2BP3, KLK5, PAR2, <t>p-AKT1</t> <t>(ser473)</t> and AKT1 in different treatment groups, GAPDH was used as the endogenous control. Unpaired t test was used in (A, C) Data is shown as mean ± SD; ** P < 0.01 and *** P < 0.001.
    P Akt1 Ser473, 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
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    p akt1 ser473 - by Bioz Stars, 2023-01
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    1) Product Images from "IGF2BP3 promotes progression of gallbladder carcinoma by stabilizing KLK5 mRNA in N 6 -methyladenosine-dependent binding"

    Article Title: IGF2BP3 promotes progression of gallbladder carcinoma by stabilizing KLK5 mRNA in N 6 -methyladenosine-dependent binding

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2022.1035871

    KLK5 rescued the abrogated GBC progression in IGF2BP3-depleted cells. (A–E) IGF2BP3-depleted GBC cells were co-transfected with PLVX-KLK5 or control plasmid. Cell growth ability was assessed by CCK8 assay (A) , colony formation assay (B) and EdU assay ( C , 100X, scale bar: 100μm). Statistical significance was analyzed based on the number of EdU stained cells (right). (D) Transwell assays were conducted to compare the migration ability in each treated group of GBC cells (100X, scale bar: 100μm). (E) Western blotting assays showed the expression of IGF2BP3, KLK5, PAR2, p-AKT1 (ser473) and AKT1 in different treatment groups, GAPDH was used as the endogenous control. Unpaired t test was used in (A, C) Data is shown as mean ± SD; ** P < 0.01 and *** P < 0.001.
    Figure Legend Snippet: KLK5 rescued the abrogated GBC progression in IGF2BP3-depleted cells. (A–E) IGF2BP3-depleted GBC cells were co-transfected with PLVX-KLK5 or control plasmid. Cell growth ability was assessed by CCK8 assay (A) , colony formation assay (B) and EdU assay ( C , 100X, scale bar: 100μm). Statistical significance was analyzed based on the number of EdU stained cells (right). (D) Transwell assays were conducted to compare the migration ability in each treated group of GBC cells (100X, scale bar: 100μm). (E) Western blotting assays showed the expression of IGF2BP3, KLK5, PAR2, p-AKT1 (ser473) and AKT1 in different treatment groups, GAPDH was used as the endogenous control. Unpaired t test was used in (A, C) Data is shown as mean ± SD; ** P < 0.01 and *** P < 0.001.

    Techniques Used: Transfection, Plasmid Preparation, CCK-8 Assay, Colony Assay, EdU Assay, Staining, Migration, Western Blot, Expressing

    pakt s308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt s308
    Antibodies used for immunohistochemistry and western blots.
    Pakt S308, 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
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    Average 94 stars, based on 1 article reviews
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    1) Product Images from "RASSF1A independence and early galectin‐1 upregulation in PIK3CA‐induced hepatocarcinogenesis: new therapeutic venues"

    Article Title: RASSF1A independence and early galectin‐1 upregulation in PIK3CA‐induced hepatocarcinogenesis: new therapeutic venues

    Journal: Molecular Oncology

    doi: 10.1002/1878-0261.13135

    Antibodies used for immunohistochemistry and western blots.
    Figure Legend Snippet: Antibodies used for immunohistochemistry and western blots.

    Techniques Used: Immunohistochemistry, Western Blot

    pakt s473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt s473
    Antibodies used for immunohistochemistry and western blots.
    Pakt S473, 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/pakt s473/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pakt s473 - by Bioz Stars, 2023-01
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    1) Product Images from "RASSF1A independence and early galectin‐1 upregulation in PIK3CA‐induced hepatocarcinogenesis: new therapeutic venues"

    Article Title: RASSF1A independence and early galectin‐1 upregulation in PIK3CA‐induced hepatocarcinogenesis: new therapeutic venues

    Journal: Molecular Oncology

    doi: 10.1002/1878-0261.13135

    Antibodies used for immunohistochemistry and western blots.
    Figure Legend Snippet: Antibodies used for immunohistochemistry and western blots.

    Techniques Used: Immunohistochemistry, Western Blot

    pakt s473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt s473
    Primary and secondary antibodies for Western Blotting
    Pakt S473, 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/pakt s473/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
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    1) Product Images from "Dexibuprofen ameliorates peripheral and central risk factors associated with Alzheimer’s disease in metabolically stressed APPswe/PS1dE9 mice"

    Article Title: Dexibuprofen ameliorates peripheral and central risk factors associated with Alzheimer’s disease in metabolically stressed APPswe/PS1dE9 mice

    Journal: Cell & Bioscience

    doi: 10.1186/s13578-021-00646-w

    Primary and secondary antibodies for Western Blotting
    Figure Legend Snippet: Primary and secondary antibodies for Western Blotting

    Techniques Used: Western Blot

    pakt s473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt s473
    Pakt S473, 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/pakt s473/product/Cell Signaling Technology Inc
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    pakt s473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt s473
    KEY RESOURCES TABLE
    Pakt S473, 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
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    1) Product Images from "Hyperactivation of TORC1 drives resistance to the pan-HER tyrosine kinase inhibitor neratinib in HER2- mutant cancers"

    Article Title: Hyperactivation of TORC1 drives resistance to the pan-HER tyrosine kinase inhibitor neratinib in HER2- mutant cancers

    Journal: Cancer cell

    doi: 10.1016/j.ccell.2019.12.013

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Amplification, Activation Assay, Sequencing, Recombinant, shRNA, Variant Assay, Software

    pakt t308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt t308
    KEY RESOURCES TABLE
    Pakt T308, 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
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    Average 94 stars, based on 1 article reviews
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    1) Product Images from "Hyperactivation of TORC1 drives resistance to the pan-HER tyrosine kinase inhibitor neratinib in HER2- mutant cancers"

    Article Title: Hyperactivation of TORC1 drives resistance to the pan-HER tyrosine kinase inhibitor neratinib in HER2- mutant cancers

    Journal: Cancer cell

    doi: 10.1016/j.ccell.2019.12.013

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Amplification, Activation Assay, Sequencing, Recombinant, shRNA, Variant Assay, Software

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    Cell Signaling Technology Inc ser473
    BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT <t>(Ser473),</t> total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.
    Ser473, 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/ser473/product/Cell Signaling Technology Inc
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    BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT <t>(Ser473),</t> total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.
    P Akt1 2 3 Ser473, 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
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    ( A ) Phalloidin detection shows cells with polarized F-actin (white arrow) following treatment with recombinant Periostin compared to vehicle control. Scale bars represent 10 μm. ( B ) Graphic represents percentage of cells with stress fiber formation (polarized F-actin) after periostin or vehicle stimuli. Results were determined by measuring fields using independent triplicates (**p<0.01) ( C ) Activation of PI3K and mTOR signaling is triggered by Periostin treatment in a dose-dependent manner, as detected by phosphorylated AKT at Threonine 308 <t>(pAKT</t> Thr308 ) and <t>Serine</t> <t>473</t> (pAKT Ser473 ) and phosphorylated S6 (pS6). Note that 50 ng/ml of Periostin is the optimal concentration for PI3K activation. GAPDH was used as a loading control.
    Pakt S473, 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
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    KLK5 rescued the abrogated GBC progression in IGF2BP3-depleted cells. (A–E) IGF2BP3-depleted GBC cells were co-transfected with PLVX-KLK5 or control plasmid. Cell growth ability was assessed by CCK8 assay (A) , colony formation assay (B) and EdU assay ( C , 100X, scale bar: 100μm). Statistical significance was analyzed based on the number of EdU stained cells (right). (D) Transwell assays were conducted to compare the migration ability in each treated group of GBC cells (100X, scale bar: 100μm). (E) Western blotting assays showed the expression of IGF2BP3, KLK5, PAR2, <t>p-AKT1</t> <t>(ser473)</t> and AKT1 in different treatment groups, GAPDH was used as the endogenous control. Unpaired t test was used in (A, C) Data is shown as mean ± SD; ** P < 0.01 and *** P < 0.001.
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    Antibodies used for immunohistochemistry and western blots.
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    KEY RESOURCES TABLE
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    Image Search Results


    BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT (Ser473), total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.

    Journal: Theranostics

    Article Title: BCAP31 drives TNBC development by modulating ligand-independent EGFR trafficking and spontaneous EGFR phosphorylation

    doi: 10.7150/thno.35383

    Figure Lengend Snippet: BCAP31 interacts with EGFR and facilitates ligand-independent EGFR/AKT pathway activation. (A) Mass spectrometry (MS) analysis of BCAP31-associated proteins. The purified protein complex was resolved on SDS-PAGE, followed by silver staining; then, the bands were retrieved and analysed by MS. (B) Analyses of the identified BCAP31 interactors. The diagram depicts BCAP31 interactors as detected by MS. The network was built based on the interaction network of EGFR-associated signalling and trafficking processes in the KEGG database overlaid with MS data. (C and D) The interaction between exogenous and endogenous BCAP31 and EGFR. Co-IP assay and immunoblot analyses evaluating the BCAP31-EGFR interaction in COS-7 cells and TNBC cells. (E) Downregulation of BCAP31 significantly attenuates ligand-independent signalling. TNBC cells stably expressed shBCAP31, control shRNA, or shRES when treated with EGF or EGF combined with gefitinib. IB examinations of phosphorylated EGFR and downstream signalling are shown. (F) Re-expression of the K721A kinase-dead (EGFR KD ) EGFR rescues the ligand-independent phosphorylation of EGFR at Tyr 845 and downstream AKT signalling in EGFR-knockdown MDA-MB-231 cells expressing control shRNA but not shBCAP31. SiRNA-resistant EGFR-KD was expressed in control or shBCAP31-MDA-MB-231 cells by lentivirus-mediated infection. Endogenous EGFR was knocked down by siRNA, and whole-cell lysate was harvested for western blot analysis. (G) Tumours of MDA-MB-231 tumour xenografts were harvested, homogenized, and analysed for pEGFR (Tyr1068, Tyr845), pAKT (Ser473), total EGFR and total AKT by ELISA. Statistical significance was determined using Dunnett's test.

    Article Snippet: All the ELISA kits were provided by Cell Signaling Technology, and the catalogue numbers are #7189 (Tyr845 of pEGFR), #7134 (Ser473 of pAKT), #7240 (Tyr1068 of pEGFR), and #7177 (Thr202/Tyr204 of pERK).

    Techniques: Activation Assay, Mass Spectrometry, Purification, SDS Page, Silver Staining, Co-Immunoprecipitation Assay, Western Blot, Stable Transfection, shRNA, Expressing, Infection, Enzyme-linked Immunosorbent Assay

    ( A ) Phalloidin detection shows cells with polarized F-actin (white arrow) following treatment with recombinant Periostin compared to vehicle control. Scale bars represent 10 μm. ( B ) Graphic represents percentage of cells with stress fiber formation (polarized F-actin) after periostin or vehicle stimuli. Results were determined by measuring fields using independent triplicates (**p<0.01) ( C ) Activation of PI3K and mTOR signaling is triggered by Periostin treatment in a dose-dependent manner, as detected by phosphorylated AKT at Threonine 308 (pAKT Thr308 ) and Serine 473 (pAKT Ser473 ) and phosphorylated S6 (pS6). Note that 50 ng/ml of Periostin is the optimal concentration for PI3K activation. GAPDH was used as a loading control.

    Journal: PLoS ONE

    Article Title: Periostin Responds to Mechanical Stress and Tension by Activating the MTOR Signaling Pathway

    doi: 10.1371/journal.pone.0083580

    Figure Lengend Snippet: ( A ) Phalloidin detection shows cells with polarized F-actin (white arrow) following treatment with recombinant Periostin compared to vehicle control. Scale bars represent 10 μm. ( B ) Graphic represents percentage of cells with stress fiber formation (polarized F-actin) after periostin or vehicle stimuli. Results were determined by measuring fields using independent triplicates (**p<0.01) ( C ) Activation of PI3K and mTOR signaling is triggered by Periostin treatment in a dose-dependent manner, as detected by phosphorylated AKT at Threonine 308 (pAKT Thr308 ) and Serine 473 (pAKT Ser473 ) and phosphorylated S6 (pS6). Note that 50 ng/ml of Periostin is the optimal concentration for PI3K activation. GAPDH was used as a loading control.

    Article Snippet: Membranes were blocked in 0.1 M Tris (pH 7.5), 0.9% NaCl and 0.05% Tween-20 (TBS-T) containing 5% nonfat dry milk and probed with anti-Periostin (Biovendor), pS6 (Dako), pAKT T308 (Abcam) and pAKT S473 (Cell Signaling) antibodies.

    Techniques: Recombinant, Activation Assay, Concentration Assay

    KLK5 rescued the abrogated GBC progression in IGF2BP3-depleted cells. (A–E) IGF2BP3-depleted GBC cells were co-transfected with PLVX-KLK5 or control plasmid. Cell growth ability was assessed by CCK8 assay (A) , colony formation assay (B) and EdU assay ( C , 100X, scale bar: 100μm). Statistical significance was analyzed based on the number of EdU stained cells (right). (D) Transwell assays were conducted to compare the migration ability in each treated group of GBC cells (100X, scale bar: 100μm). (E) Western blotting assays showed the expression of IGF2BP3, KLK5, PAR2, p-AKT1 (ser473) and AKT1 in different treatment groups, GAPDH was used as the endogenous control. Unpaired t test was used in (A, C) Data is shown as mean ± SD; ** P < 0.01 and *** P < 0.001.

    Journal: Frontiers in Oncology

    Article Title: IGF2BP3 promotes progression of gallbladder carcinoma by stabilizing KLK5 mRNA in N 6 -methyladenosine-dependent binding

    doi: 10.3389/fonc.2022.1035871

    Figure Lengend Snippet: KLK5 rescued the abrogated GBC progression in IGF2BP3-depleted cells. (A–E) IGF2BP3-depleted GBC cells were co-transfected with PLVX-KLK5 or control plasmid. Cell growth ability was assessed by CCK8 assay (A) , colony formation assay (B) and EdU assay ( C , 100X, scale bar: 100μm). Statistical significance was analyzed based on the number of EdU stained cells (right). (D) Transwell assays were conducted to compare the migration ability in each treated group of GBC cells (100X, scale bar: 100μm). (E) Western blotting assays showed the expression of IGF2BP3, KLK5, PAR2, p-AKT1 (ser473) and AKT1 in different treatment groups, GAPDH was used as the endogenous control. Unpaired t test was used in (A, C) Data is shown as mean ± SD; ** P < 0.01 and *** P < 0.001.

    Article Snippet: The targeted proteins were detected using different antibodies, including GAPDH (1:1000, Proteintech, USA), IGF2BP3 (1:1000, Proteintech, USA), AKT1 (1:1000, Cell Signaling Technology, USA), P-AKT1(ser473) (1:1000, Cell Signaling Technology, USA), KLK5 (1:1000, #abs136657, Abisin, Shanghai), PAR2 (1:1000, Abclonal Technology Co., Ltd.) and METTL3 (1:1000, Abclonal Technology Co., Ltd.).

    Techniques: Transfection, Plasmid Preparation, CCK-8 Assay, Colony Assay, EdU Assay, Staining, Migration, Western Blot, Expressing

    Antibodies used for immunohistochemistry and western blots.

    Journal: Molecular Oncology

    Article Title: RASSF1A independence and early galectin‐1 upregulation in PIK3CA‐induced hepatocarcinogenesis: new therapeutic venues

    doi: 10.1002/1878-0261.13135

    Figure Lengend Snippet: Antibodies used for immunohistochemistry and western blots.

    Article Snippet: pAKT (S308) , 1 : 1000 , Cell Signaling Technology, Inc. , 13038.

    Techniques: Immunohistochemistry, Western Blot

    KEY RESOURCES TABLE

    Journal: Cancer cell

    Article Title: Hyperactivation of TORC1 drives resistance to the pan-HER tyrosine kinase inhibitor neratinib in HER2- mutant cancers

    doi: 10.1016/j.ccell.2019.12.013

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: pAKT (T308) , Cell Signaling Technologies , Cat# 13038.

    Techniques: Amplification, Activation Assay, Sequencing, Recombinant, shRNA, Variant Assay, Software