phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab
    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho akt ser473 d9e xp rabbit mab/product/Cell Signaling Technology Inc
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    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab

    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho akt ser473 d9e xp rabbit mab/product/Cell Signaling Technology Inc
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    1) Product Images from "Enniatin A inhibits the chaperone Hsp90 and unleashes the immune system against triple-negative breast cancer"

    Article Title: Enniatin A inhibits the chaperone Hsp90 and unleashes the immune system against triple-negative breast cancer

    Journal: iScience

    doi: 10.1016/j.isci.2023.108308


    Figure Legend Snippet:

    Techniques Used: Affinity Purification, Purification, Recombinant, Proliferation Assay, Software

    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab
    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab
    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab

    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "High NEK2 expression in myeloid progenitors suppresses T cell immunity in multiple myeloma"

    Article Title: High NEK2 expression in myeloid progenitors suppresses T cell immunity in multiple myeloma

    Journal: Cell Reports Medicine

    doi: 10.1016/j.xcrm.2023.101214


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    Techniques Used: Purification, Recombinant, Western Blot, Stripping, Cell Stimulation, Staining, Cell Isolation, Activation Assay, Extraction, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, Quantitation Assay, SYBR Green Assay, RNA Sequencing Assay, Mutagenesis, Knock-Out, Software

    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab
    Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at <t>Ser473</t> and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.
    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho akt ser473 d9e xp rabbit mab/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    phospho akt ser473 d9e xp rabbit mab - by Bioz Stars, 2023-11
    86/100 stars

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    1) Product Images from "Human cytomegalovirus attenuates AKT activity by destabilizing insulin receptor substrate proteins"

    Article Title: Human cytomegalovirus attenuates AKT activity by destabilizing insulin receptor substrate proteins

    Journal: Journal of Virology

    doi: 10.1128/jvi.00563-23

    Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at Ser473 and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.
    Figure Legend Snippet: Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at Ser473 and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.

    Techniques Used: Activation Assay, Scaffolding, Activity Assay, Generated, Lysis, SDS Page, Western Blot, Infection, Labeling, Staining

    Antibodies used in this study a
    Figure Legend Snippet: Antibodies used in this study a

    Techniques Used:

    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab
    Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at <t>Ser473</t> and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.
    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho akt ser473 d9e xp rabbit mab/product/Cell Signaling Technology Inc
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    phospho akt ser473 d9e xp rabbit mab - by Bioz Stars, 2023-11
    86/100 stars

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    1) Product Images from "Human cytomegalovirus attenuates AKT activity by destabilizing insulin receptor substrate proteins"

    Article Title: Human cytomegalovirus attenuates AKT activity by destabilizing insulin receptor substrate proteins

    Journal: Journal of Virology

    doi: 10.1128/jvi.00563-23

    Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at Ser473 and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.
    Figure Legend Snippet: Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at Ser473 and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.

    Techniques Used: Activation Assay, Scaffolding, Activity Assay, Generated, Lysis, SDS Page, Western Blot, Infection, Labeling, Staining

    Antibodies used in this study a
    Figure Legend Snippet: Antibodies used in this study a

    Techniques Used:

    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab
    Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at <t>Ser473</t> and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.
    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho akt ser473 d9e xp rabbit mab/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho akt ser473 d9e xp rabbit mab - by Bioz Stars, 2023-11
    86/100 stars

    Images

    1) Product Images from "Human cytomegalovirus attenuates AKT activity by destabilizing insulin receptor substrate proteins"

    Article Title: Human cytomegalovirus attenuates AKT activity by destabilizing insulin receptor substrate proteins

    Journal: Journal of Virology

    doi: 10.1128/jvi.00563-23

    Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at Ser473 and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.
    Figure Legend Snippet: Human cytomegalovirus (HCMV) prevents AKT activation in response to serum. (A) Stimulation of various receptor tyrosine kinases (RTKs) and other growth factor receptors causes the recruitment of the scaffolding adaptor insulin receptor substrate 1 (IRS1) to the receptor complex. IRS1 is then phosphorylated, which provides a docking site for recruitment of class IA phosphoinositol 3-kinases (PI3K). PI3K phosphorylates the lipid substrate phosphoinositol 4,5 diphosphate (PIP2) to generate phosphoinositol 3,4,5 triphosphate (PIP3). The pleckstrin homology (PH) domain of AKT binds to PIP3, which drives its localization to membranes where mTORC2 and phosphoinositol-dependent kinase 1 (PDK1) are available to phosphorylate AKT at Ser473 and Thr308, respectively, activating its kinase activity. Once activated, AKT phosphorylates many substrates, such as the proline-rich AKT substrate of 40 kDa (PRAS40) at Thr246, Forkhead box class O (FoxO) family transcription factors, and the tuberous sclerosis complex 2 (TSC2) protei. Phosphorylation of TSC2 leads to activation of mTORC1, which in turn phosphorylates and activates S6 kinase (S6K). S6K phosphorylates S6 to increase protein synthesis. (Schematic generated in BioRender.) (B) Serum-starved fibroblasts were stimulated with insulin or 5% newborn calf serum for the indicated times (min) prior to lysis. Protein extracts were then resolved by SDS-PAGE, and transferred to nitrocellulose membranes for Western blot analysis using a phospho-specific antibody to detect AKT phosphorylated at Ser473 compared with AKT (pan) antibody. (C–G) Fibroblasts were serum starved overnight and either mock infected or infected with HCMV strain Towne at multiplicity of infection (MOI) of 2 (MOI = 2 TCID50/cell) for the indicated times, hours post-infection (hpi). Cells were then either treated (or mock treated) with serum for 10 min prior to lysis for Western blot analysis. (E–G) Fluorescent signals from dye-labeled secondary antibodies were quantified, and phospho-specific antibody detection of pAKT Thr308, pAKT Ser473, and PRAS40 Thr246 was normalized to AKT (pan) or PRAS40. The arithmetic mean was calculated and graphed. Error bars indicate standard error of the mean (SEM) from three independent biological replicates. Where indicated, Ponceau staining was used to visualize protein loading and confirm efficient transfer.

    Techniques Used: Activation Assay, Scaffolding, Activity Assay, Generated, Lysis, SDS Page, Western Blot, Infection, Labeling, Staining

    Antibodies used in this study a
    Figure Legend Snippet: Antibodies used in this study a

    Techniques Used:

    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    • 86
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    Structured Review

    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab
    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho akt ser473 d9e xp rabbit mab/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho akt ser473 d9e xp rabbit mab - by Bioz Stars, 2023-11
    86/100 stars

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    • phospho akt ser473 d9e xp rabbit mab  (Cell Signaling Technology Inc)
    86
    Cell Signaling Technology Inc phospho akt ser473 d9e xp rabbit mab
    Phospho Akt Ser473 D9e Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho akt ser473 d9e xp rabbit mab/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho akt ser473 d9e xp rabbit mab - by Bioz Stars, 2023-11
    86/100 stars
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