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pcdna3.1(+) myc-akap2 dsg1  (GenScript corporation)

 
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    Structured Review

    GenScript corporation pcdna3.1(+) myc-akap2 dsg1
    (A) Schematic depicting <t>AKAP2</t> showing the PKA binding domain and relative positions of the three putative βTrCP binding sites in AKAP2. The corresponding amino acids for each motif are 382–384 <t>(DSG1),</t> 472–474 (DSG2), and 502–504 (DSG3). (B) Sequence alignment of the DSG2 motif in Homo sapiens , Mus musculus , and Xenopus laevis. The CDC25B degron sequence for βTrCP is also shown, demonstrating a known example that is divergent from the canonical DSGxxS sequence. The sequence of the AKAP2 DSG2-motif mutant used in future experiments is shown below. (C) HEK293T cells were co-transfected with plasmids expressing Myc-AKAP2 (WT or DSG degron mutants) together with an empty vector control or FLAG-βTrCP2. Twenty-four hours post-transfection, the cells were collected, and βTrCP was immunoprecipitated using anti-FLAG affinity gel. Eluates were analyzed by immunoblot for the indicated proteins. n = 3. (D) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were treated with 100 ng/mL nocodazole for 16 h to synchronize cells in mitosis. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (E) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were co-treated with 100 ng/mL nocodazole plus DMSO or 100 nM BI2536 for 16 h. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (F) HEK293T cells were co-transfected with the indicated plasmids for 48 h. After 48 h, the cells were collected and lysed under denaturing conditions. Ubiquitinated proteins were captured using Ni-NTA pull-down. The ubiquitination status of AKAP2 was analyzed by immunoblot. Cells were treated with 20 μM MG132 and 20 μM PR619 for 4 h prior to collection. n = 2. (G) In vitro ubiquitination reactions using TAMRA-labeled AKAP2 WT peptide or AKAP2 phospho-peptide (p-S473) as a substrate, monitored by fluorescence scanning of an SDS-PAGE gel. Representative of n = 4 independent experiments.
    Pcdna3.1(+) Myc Akap2 Dsg1, supplied by GenScript corporation, 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/pcdna3.1(+) myc-akap2 dsg1/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    pcdna3.1(+) myc-akap2 dsg1 - by Bioz Stars, 2026-02
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    1) Product Images from "Proteomic analysis reveals a PLK1-dependent G2/M degradation program and a role for AKAP2 in coordinating the mitotic cytoskeleton"

    Article Title: Proteomic analysis reveals a PLK1-dependent G2/M degradation program and a role for AKAP2 in coordinating the mitotic cytoskeleton

    Journal: Cell reports

    doi: 10.1016/j.celrep.2024.114510

    (A) Schematic depicting AKAP2 showing the PKA binding domain and relative positions of the three putative βTrCP binding sites in AKAP2. The corresponding amino acids for each motif are 382–384 (DSG1), 472–474 (DSG2), and 502–504 (DSG3). (B) Sequence alignment of the DSG2 motif in Homo sapiens , Mus musculus , and Xenopus laevis. The CDC25B degron sequence for βTrCP is also shown, demonstrating a known example that is divergent from the canonical DSGxxS sequence. The sequence of the AKAP2 DSG2-motif mutant used in future experiments is shown below. (C) HEK293T cells were co-transfected with plasmids expressing Myc-AKAP2 (WT or DSG degron mutants) together with an empty vector control or FLAG-βTrCP2. Twenty-four hours post-transfection, the cells were collected, and βTrCP was immunoprecipitated using anti-FLAG affinity gel. Eluates were analyzed by immunoblot for the indicated proteins. n = 3. (D) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were treated with 100 ng/mL nocodazole for 16 h to synchronize cells in mitosis. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (E) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were co-treated with 100 ng/mL nocodazole plus DMSO or 100 nM BI2536 for 16 h. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (F) HEK293T cells were co-transfected with the indicated plasmids for 48 h. After 48 h, the cells were collected and lysed under denaturing conditions. Ubiquitinated proteins were captured using Ni-NTA pull-down. The ubiquitination status of AKAP2 was analyzed by immunoblot. Cells were treated with 20 μM MG132 and 20 μM PR619 for 4 h prior to collection. n = 2. (G) In vitro ubiquitination reactions using TAMRA-labeled AKAP2 WT peptide or AKAP2 phospho-peptide (p-S473) as a substrate, monitored by fluorescence scanning of an SDS-PAGE gel. Representative of n = 4 independent experiments.
    Figure Legend Snippet: (A) Schematic depicting AKAP2 showing the PKA binding domain and relative positions of the three putative βTrCP binding sites in AKAP2. The corresponding amino acids for each motif are 382–384 (DSG1), 472–474 (DSG2), and 502–504 (DSG3). (B) Sequence alignment of the DSG2 motif in Homo sapiens , Mus musculus , and Xenopus laevis. The CDC25B degron sequence for βTrCP is also shown, demonstrating a known example that is divergent from the canonical DSGxxS sequence. The sequence of the AKAP2 DSG2-motif mutant used in future experiments is shown below. (C) HEK293T cells were co-transfected with plasmids expressing Myc-AKAP2 (WT or DSG degron mutants) together with an empty vector control or FLAG-βTrCP2. Twenty-four hours post-transfection, the cells were collected, and βTrCP was immunoprecipitated using anti-FLAG affinity gel. Eluates were analyzed by immunoblot for the indicated proteins. n = 3. (D) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were treated with 100 ng/mL nocodazole for 16 h to synchronize cells in mitosis. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (E) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were co-treated with 100 ng/mL nocodazole plus DMSO or 100 nM BI2536 for 16 h. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (F) HEK293T cells were co-transfected with the indicated plasmids for 48 h. After 48 h, the cells were collected and lysed under denaturing conditions. Ubiquitinated proteins were captured using Ni-NTA pull-down. The ubiquitination status of AKAP2 was analyzed by immunoblot. Cells were treated with 20 μM MG132 and 20 μM PR619 for 4 h prior to collection. n = 2. (G) In vitro ubiquitination reactions using TAMRA-labeled AKAP2 WT peptide or AKAP2 phospho-peptide (p-S473) as a substrate, monitored by fluorescence scanning of an SDS-PAGE gel. Representative of n = 4 independent experiments.

    Techniques Used: Binding Assay, Sequencing, Mutagenesis, Transfection, Expressing, Plasmid Preparation, Control, Immunoprecipitation, Western Blot, Stable Transfection, Ubiquitin Proteomics, In Vitro, Labeling, Fluorescence, SDS Page


    Figure Legend Snippet:

    Techniques Used: Virus, Recombinant, Infection, Transfection, Western Blot, Saline, Sequencing, Plasmid Preparation, Mutagenesis, Bicinchoninic Acid Protein Assay, Staining, Expressing, Luciferase, Software, Mass Spectrometry



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    pcdna3.1(+) myc-akap2 dsg1  (GenScript corporation)
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    GenScript corporation pcdna3.1(+) myc-akap2 dsg1
    (A) Schematic depicting <t>AKAP2</t> showing the PKA binding domain and relative positions of the three putative βTrCP binding sites in AKAP2. The corresponding amino acids for each motif are 382–384 <t>(DSG1),</t> 472–474 (DSG2), and 502–504 (DSG3). (B) Sequence alignment of the DSG2 motif in Homo sapiens , Mus musculus , and Xenopus laevis. The CDC25B degron sequence for βTrCP is also shown, demonstrating a known example that is divergent from the canonical DSGxxS sequence. The sequence of the AKAP2 DSG2-motif mutant used in future experiments is shown below. (C) HEK293T cells were co-transfected with plasmids expressing Myc-AKAP2 (WT or DSG degron mutants) together with an empty vector control or FLAG-βTrCP2. Twenty-four hours post-transfection, the cells were collected, and βTrCP was immunoprecipitated using anti-FLAG affinity gel. Eluates were analyzed by immunoblot for the indicated proteins. n = 3. (D) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were treated with 100 ng/mL nocodazole for 16 h to synchronize cells in mitosis. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (E) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were co-treated with 100 ng/mL nocodazole plus DMSO or 100 nM BI2536 for 16 h. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (F) HEK293T cells were co-transfected with the indicated plasmids for 48 h. After 48 h, the cells were collected and lysed under denaturing conditions. Ubiquitinated proteins were captured using Ni-NTA pull-down. The ubiquitination status of AKAP2 was analyzed by immunoblot. Cells were treated with 20 μM MG132 and 20 μM PR619 for 4 h prior to collection. n = 2. (G) In vitro ubiquitination reactions using TAMRA-labeled AKAP2 WT peptide or AKAP2 phospho-peptide (p-S473) as a substrate, monitored by fluorescence scanning of an SDS-PAGE gel. Representative of n = 4 independent experiments.
    Pcdna3.1(+) Myc Akap2 Dsg1, supplied by GenScript corporation, 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/pcdna3.1(+) myc-akap2 dsg1/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    pcdna3.1(+) myc-akap2 dsg1 - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) Schematic depicting AKAP2 showing the PKA binding domain and relative positions of the three putative βTrCP binding sites in AKAP2. The corresponding amino acids for each motif are 382–384 (DSG1), 472–474 (DSG2), and 502–504 (DSG3). (B) Sequence alignment of the DSG2 motif in Homo sapiens , Mus musculus , and Xenopus laevis. The CDC25B degron sequence for βTrCP is also shown, demonstrating a known example that is divergent from the canonical DSGxxS sequence. The sequence of the AKAP2 DSG2-motif mutant used in future experiments is shown below. (C) HEK293T cells were co-transfected with plasmids expressing Myc-AKAP2 (WT or DSG degron mutants) together with an empty vector control or FLAG-βTrCP2. Twenty-four hours post-transfection, the cells were collected, and βTrCP was immunoprecipitated using anti-FLAG affinity gel. Eluates were analyzed by immunoblot for the indicated proteins. n = 3. (D) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were treated with 100 ng/mL nocodazole for 16 h to synchronize cells in mitosis. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (E) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were co-treated with 100 ng/mL nocodazole plus DMSO or 100 nM BI2536 for 16 h. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (F) HEK293T cells were co-transfected with the indicated plasmids for 48 h. After 48 h, the cells were collected and lysed under denaturing conditions. Ubiquitinated proteins were captured using Ni-NTA pull-down. The ubiquitination status of AKAP2 was analyzed by immunoblot. Cells were treated with 20 μM MG132 and 20 μM PR619 for 4 h prior to collection. n = 2. (G) In vitro ubiquitination reactions using TAMRA-labeled AKAP2 WT peptide or AKAP2 phospho-peptide (p-S473) as a substrate, monitored by fluorescence scanning of an SDS-PAGE gel. Representative of n = 4 independent experiments.

    Journal: Cell reports

    Article Title: Proteomic analysis reveals a PLK1-dependent G2/M degradation program and a role for AKAP2 in coordinating the mitotic cytoskeleton

    doi: 10.1016/j.celrep.2024.114510

    Figure Lengend Snippet: (A) Schematic depicting AKAP2 showing the PKA binding domain and relative positions of the three putative βTrCP binding sites in AKAP2. The corresponding amino acids for each motif are 382–384 (DSG1), 472–474 (DSG2), and 502–504 (DSG3). (B) Sequence alignment of the DSG2 motif in Homo sapiens , Mus musculus , and Xenopus laevis. The CDC25B degron sequence for βTrCP is also shown, demonstrating a known example that is divergent from the canonical DSGxxS sequence. The sequence of the AKAP2 DSG2-motif mutant used in future experiments is shown below. (C) HEK293T cells were co-transfected with plasmids expressing Myc-AKAP2 (WT or DSG degron mutants) together with an empty vector control or FLAG-βTrCP2. Twenty-four hours post-transfection, the cells were collected, and βTrCP was immunoprecipitated using anti-FLAG affinity gel. Eluates were analyzed by immunoblot for the indicated proteins. n = 3. (D) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were treated with 100 ng/mL nocodazole for 16 h to synchronize cells in mitosis. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (E) HCT116 cells stably expressing FLAG-AKAP2 WT or FLAG-AKAP2 DSG2 were grown asynchronously or were co-treated with 100 ng/mL nocodazole plus DMSO or 100 nM BI2536 for 16 h. After 16 h, mitotic-arrested cells were collected using the mitotic shake-off procedure. All cell lysates were analyzed by immunoblot for the indicated proteins. n = 3. (F) HEK293T cells were co-transfected with the indicated plasmids for 48 h. After 48 h, the cells were collected and lysed under denaturing conditions. Ubiquitinated proteins were captured using Ni-NTA pull-down. The ubiquitination status of AKAP2 was analyzed by immunoblot. Cells were treated with 20 μM MG132 and 20 μM PR619 for 4 h prior to collection. n = 2. (G) In vitro ubiquitination reactions using TAMRA-labeled AKAP2 WT peptide or AKAP2 phospho-peptide (p-S473) as a substrate, monitored by fluorescence scanning of an SDS-PAGE gel. Representative of n = 4 independent experiments.

    Article Snippet: pcDNA3.1(+) Myc-AKAP2 DSG1 , GenScript , N/A.

    Techniques: Binding Assay, Sequencing, Mutagenesis, Transfection, Expressing, Plasmid Preparation, Control, Immunoprecipitation, Western Blot, Stable Transfection, Ubiquitin Proteomics, In Vitro, Labeling, Fluorescence, SDS Page

    Journal: Cell reports

    Article Title: Proteomic analysis reveals a PLK1-dependent G2/M degradation program and a role for AKAP2 in coordinating the mitotic cytoskeleton

    doi: 10.1016/j.celrep.2024.114510

    Figure Lengend Snippet:

    Article Snippet: pcDNA3.1(+) Myc-AKAP2 DSG1 , GenScript , N/A.

    Techniques: Virus, Recombinant, Infection, Transfection, Western Blot, Saline, Sequencing, Plasmid Preparation, Mutagenesis, Bicinchoninic Acid Protein Assay, Staining, Expressing, Luciferase, Software, Mass Spectrometry