p akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p akt308
    P Akt308, 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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    p akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p akt308
    Effects of deoxyshikonin on Akt/PI3K/mTOR signalling pathway in HT29 cells. (A) PI3K, p -PI3K, Akt, p <t>-Akt308</t> and mTOR proteins in HT29 and DLD-1 cell lines were determined with specific antibodies. β-Actin was used as loading control. The relative intensity of each protein was normalized with β-actin. (B) The expression levels of Akt, PI3K, mTOR mRNA in HT29 and DLD-1 cell lines were measured by RT-PCR. Data were results of three independent experiments with mean ± SD. * p < 0.05, ** p < 0.001, *** p < 0.0001 and **** p < 0.00001 versus control (0 μg/mL).
    P Akt308, 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 "Deoxyshikonin isolated from Arnebia euchroma inhibits colorectal cancer by down-regulating the PI3K/Akt/mTOR pathway"

    Article Title: Deoxyshikonin isolated from Arnebia euchroma inhibits colorectal cancer by down-regulating the PI3K/Akt/mTOR pathway

    Journal: Pharmaceutical Biology

    doi: 10.1080/13880209.2019.1626447

    Effects of deoxyshikonin on Akt/PI3K/mTOR signalling pathway in HT29 cells. (A) PI3K, p -PI3K, Akt, p -Akt308 and mTOR proteins in HT29 and DLD-1 cell lines were determined with specific antibodies. β-Actin was used as loading control. The relative intensity of each protein was normalized with β-actin. (B) The expression levels of Akt, PI3K, mTOR mRNA in HT29 and DLD-1 cell lines were measured by RT-PCR. Data were results of three independent experiments with mean ± SD. * p < 0.05, ** p < 0.001, *** p < 0.0001 and **** p < 0.00001 versus control (0 μg/mL).
    Figure Legend Snippet: Effects of deoxyshikonin on Akt/PI3K/mTOR signalling pathway in HT29 cells. (A) PI3K, p -PI3K, Akt, p -Akt308 and mTOR proteins in HT29 and DLD-1 cell lines were determined with specific antibodies. β-Actin was used as loading control. The relative intensity of each protein was normalized with β-actin. (B) The expression levels of Akt, PI3K, mTOR mRNA in HT29 and DLD-1 cell lines were measured by RT-PCR. Data were results of three independent experiments with mean ± SD. * p < 0.05, ** p < 0.001, *** p < 0.0001 and **** p < 0.00001 versus control (0 μg/mL).

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    Deoxyshikonin mediated HT29 cells viability, cell apoptosis and cycle arrest depended on the PI3K/Akt/mTOR signalling pathway. HT29 cells were pretreated with LY294002, or NVP-BEZ235, or MK-2206, then co-treated with 50 μg/mL deoxyshikonin for further 48 h. (A) Cell viability was assessed by MTT assay; (B) cell apoptosis and cell cycle arrest were detected by flow cytometry; (C) Akt and p -Akt308 proteins were determined by Western blotting. Data were presented as mean ± SD of three independent tests. * p < 0.05, ** p < 0.001, *** p < 0.0001 and **** p < 0.00001 versus control (0 μg/mL).
    Figure Legend Snippet: Deoxyshikonin mediated HT29 cells viability, cell apoptosis and cycle arrest depended on the PI3K/Akt/mTOR signalling pathway. HT29 cells were pretreated with LY294002, or NVP-BEZ235, or MK-2206, then co-treated with 50 μg/mL deoxyshikonin for further 48 h. (A) Cell viability was assessed by MTT assay; (B) cell apoptosis and cell cycle arrest were detected by flow cytometry; (C) Akt and p -Akt308 proteins were determined by Western blotting. Data were presented as mean ± SD of three independent tests. * p < 0.05, ** p < 0.001, *** p < 0.0001 and **** p < 0.00001 versus control (0 μg/mL).

    Techniques Used: MTT Assay, Flow Cytometry, Western Blot

    Anticancer activity of deoxyshikonin in vivo via PI3K/Akt/mTOR pathway. BALB/c nude mice were injected with DLD-1 cells into the subcutaneous tissue of the right auxiliary region. Xenograft model was established when tumours reached an average size of 62.5 mm 3 , and the treatments were given different drugs by intraperitoneal injection for a total of 13 days: control, DMSO (1% DMSO, every two days), deoxyshikonin (20 mg/kg in 1% DMSO, every two days), irinotecan (66.7 mg/kg, every four days). (A) Body weights and tumour volumes were measured once daily; (B) images, tumour weights and H&E staining (200× magnification) were measured after 13 days of treatment; (C) the proteins of tumour tissues were blotted with antibodies against indicated proteins (PI3K, p-PI3K, Akt, p-Akt308 and mTOR), and β-actin was blotted as a loading control. Data are presented as mean ± SD. * p < 0.05, ** p < 0.001, *** p < 0.0001 and **** p < 0.00001 versus control.
    Figure Legend Snippet: Anticancer activity of deoxyshikonin in vivo via PI3K/Akt/mTOR pathway. BALB/c nude mice were injected with DLD-1 cells into the subcutaneous tissue of the right auxiliary region. Xenograft model was established when tumours reached an average size of 62.5 mm 3 , and the treatments were given different drugs by intraperitoneal injection for a total of 13 days: control, DMSO (1% DMSO, every two days), deoxyshikonin (20 mg/kg in 1% DMSO, every two days), irinotecan (66.7 mg/kg, every four days). (A) Body weights and tumour volumes were measured once daily; (B) images, tumour weights and H&E staining (200× magnification) were measured after 13 days of treatment; (C) the proteins of tumour tissues were blotted with antibodies against indicated proteins (PI3K, p-PI3K, Akt, p-Akt308 and mTOR), and β-actin was blotted as a loading control. Data are presented as mean ± SD. * p < 0.05, ** p < 0.001, *** p < 0.0001 and **** p < 0.00001 versus control.

    Techniques Used: Activity Assay, In Vivo, Injection, Staining

    p akt308 473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p akt308 473
    P Akt308 473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti p akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p akt308
    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of <t>p-Akt308</t> and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.
    Anti P Akt308, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "ACE2 pathway regulates thermogenesis and energy metabolism"

    Article Title: ACE2 pathway regulates thermogenesis and energy metabolism

    Journal: eLife

    doi: 10.7554/eLife.72266

    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of p-Akt308 and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.
    Figure Legend Snippet: Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of p-Akt308 and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.

    Techniques Used: Isolation, Cell Culture, Western Blot, Activation Assay


    Figure Legend Snippet:

    Techniques Used: Transfection, Construct, Sequencing, Software

    anti p akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p akt308
    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of <t>p-Akt308</t> and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.
    Anti P Akt308, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p akt308/product/Cell Signaling Technology Inc
    Average 96 stars, based on 1 article reviews
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    1) Product Images from "ACE2 pathway regulates thermogenesis and energy metabolism"

    Article Title: ACE2 pathway regulates thermogenesis and energy metabolism

    Journal: eLife

    doi: 10.7554/eLife.72266

    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of p-Akt308 and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.
    Figure Legend Snippet: Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of p-Akt308 and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.

    Techniques Used: Isolation, Cell Culture, Western Blot, Activation Assay


    Figure Legend Snippet:

    Techniques Used: Transfection, Construct, Sequencing, Software

    p akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p akt308
    P Akt308, 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/p akt308/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    anti p akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p akt308
    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 −6 M) for 24 hours, Akt inhibitor MK2206 (30μM) for 24 hours, PKA inhibitor H89 (30μM) for 2 hours, or adenylylcyclase inhibitor SQ-22536 (10μM) for 24 hours. (A, B) Representative western blots showing the changes of <t>p-Akt308</t> and Akt in BAT of ACE2 -/y ( A ) and db/db+ACE2 mice ( B ) exposure at 4°C. n =3/each group. (C) Representative western blots showing the Akt, uncoupling protein-1 (UCP1) and forkhead box protein O1 (FOXO1) changes. n=3/each group. (D) Relative mRNA levels of thermogenic and mitochondrial genes. n=4-6/each group. (E) Continuous measurement of oxygen consumption rate (OCR). Oxygen consumption was performed under basal conditions, following the addition of oligomycin (14μM), the pharmacological uncoupler FCCP (10μM) or the Complex III and I inhibitor antimycin A and rotenone (4μM each). n =5-7/each group. (F, G) Representative western blots showing the p-PKA and PKA changes in BAT of ACE2 -/y ( F ) and db/db+ACE2 mice ( G ) exposure at 4°C. n =3/each group. (H) Relative mRNA levels of thermogenic and mitochondrial genes. n=4-6/each group. (I) Representative western blots showing the UCP1 and PGC-1α changes. n=4/each group. (J) Continuous measurement of OCR. Oxygen consumption was performed under basal conditions, following the addition of oligomycin, the pharmacological uncoupler FCCP (10μM) or the Complex III and I inhibitor antimycin A and rotenone (4μM each). N =5-7/each group. Data represent mean ± SEM; * p < 0.05, ** p < 0.01 vs control group by Student’s test. # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by Student’s t -test. (K) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function. The online version of this article includes the following figure supplement(s) for : . RNA-Seq analysis of primary brown adipocytes from ACE2 -/y and WT mice; Ang-(1-7) regulates thermogenesis through Akt and PKA signaling in BAT. . Ang-(1-7) regulates thermogenesis through Akt and PKA signaling in BAT.
    Anti P Akt308, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p akt308/product/Cell Signaling Technology Inc
    Average 96 stars, based on 1 article reviews
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    anti p akt308 - by Bioz Stars, 2024-06
    96/100 stars

    Images

    1) Product Images from "ACE2 Pathway Regulates Thermogenesis and Energy Metabolism"

    Article Title: ACE2 Pathway Regulates Thermogenesis and Energy Metabolism

    Journal: bioRxiv

    doi: 10.1101/2021.08.10.455823

    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 −6 M) for 24 hours, Akt inhibitor MK2206 (30μM) for 24 hours, PKA inhibitor H89 (30μM) for 2 hours, or adenylylcyclase inhibitor SQ-22536 (10μM) for 24 hours. (A, B) Representative western blots showing the changes of p-Akt308 and Akt in BAT of ACE2 -/y ( A ) and db/db+ACE2 mice ( B ) exposure at 4°C. n =3/each group. (C) Representative western blots showing the Akt, uncoupling protein-1 (UCP1) and forkhead box protein O1 (FOXO1) changes. n=3/each group. (D) Relative mRNA levels of thermogenic and mitochondrial genes. n=4-6/each group. (E) Continuous measurement of oxygen consumption rate (OCR). Oxygen consumption was performed under basal conditions, following the addition of oligomycin (14μM), the pharmacological uncoupler FCCP (10μM) or the Complex III and I inhibitor antimycin A and rotenone (4μM each). n =5-7/each group. (F, G) Representative western blots showing the p-PKA and PKA changes in BAT of ACE2 -/y ( F ) and db/db+ACE2 mice ( G ) exposure at 4°C. n =3/each group. (H) Relative mRNA levels of thermogenic and mitochondrial genes. n=4-6/each group. (I) Representative western blots showing the UCP1 and PGC-1α changes. n=4/each group. (J) Continuous measurement of OCR. Oxygen consumption was performed under basal conditions, following the addition of oligomycin, the pharmacological uncoupler FCCP (10μM) or the Complex III and I inhibitor antimycin A and rotenone (4μM each). N =5-7/each group. Data represent mean ± SEM; * p < 0.05, ** p < 0.01 vs control group by Student’s test. # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by Student’s t -test. (K) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function. The online version of this article includes the following figure supplement(s) for : . RNA-Seq analysis of primary brown adipocytes from ACE2 -/y and WT mice; Ang-(1-7) regulates thermogenesis through Akt and PKA signaling in BAT. . Ang-(1-7) regulates thermogenesis through Akt and PKA signaling in BAT.
    Figure Legend Snippet: Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 −6 M) for 24 hours, Akt inhibitor MK2206 (30μM) for 24 hours, PKA inhibitor H89 (30μM) for 2 hours, or adenylylcyclase inhibitor SQ-22536 (10μM) for 24 hours. (A, B) Representative western blots showing the changes of p-Akt308 and Akt in BAT of ACE2 -/y ( A ) and db/db+ACE2 mice ( B ) exposure at 4°C. n =3/each group. (C) Representative western blots showing the Akt, uncoupling protein-1 (UCP1) and forkhead box protein O1 (FOXO1) changes. n=3/each group. (D) Relative mRNA levels of thermogenic and mitochondrial genes. n=4-6/each group. (E) Continuous measurement of oxygen consumption rate (OCR). Oxygen consumption was performed under basal conditions, following the addition of oligomycin (14μM), the pharmacological uncoupler FCCP (10μM) or the Complex III and I inhibitor antimycin A and rotenone (4μM each). n =5-7/each group. (F, G) Representative western blots showing the p-PKA and PKA changes in BAT of ACE2 -/y ( F ) and db/db+ACE2 mice ( G ) exposure at 4°C. n =3/each group. (H) Relative mRNA levels of thermogenic and mitochondrial genes. n=4-6/each group. (I) Representative western blots showing the UCP1 and PGC-1α changes. n=4/each group. (J) Continuous measurement of OCR. Oxygen consumption was performed under basal conditions, following the addition of oligomycin, the pharmacological uncoupler FCCP (10μM) or the Complex III and I inhibitor antimycin A and rotenone (4μM each). N =5-7/each group. Data represent mean ± SEM; * p < 0.05, ** p < 0.01 vs control group by Student’s test. # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by Student’s t -test. (K) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function. The online version of this article includes the following figure supplement(s) for : . RNA-Seq analysis of primary brown adipocytes from ACE2 -/y and WT mice; Ang-(1-7) regulates thermogenesis through Akt and PKA signaling in BAT. . Ang-(1-7) regulates thermogenesis through Akt and PKA signaling in BAT.

    Techniques Used: Isolation, Cell Culture, Western Blot, Activation Assay, RNA Sequencing Assay

    anti p akt308 4056 1 1000  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p akt308 4056 1 1000
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    western blot primary antibodies against p akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc western blot primary antibodies against p akt308
    Western Blot Primary Antibodies Against P Akt308, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p akt308
    Knockdown of ET-1 blocked the PI3K/Akt pathway. Western blot was performed to detect the level of p-PI3K, PI3K, p-Akt473, <t>p-Akt308</t> and Akt in ECA109 cells. ***, P<0.001, compared with the control group; ### , P<0.001, compared with the si-ET-1 group.
    P Akt308, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "ET-1 promotes the growth and metastasis of esophageal squamous cell carcinoma via activating PI3K/Akt pathway"

    Article Title: ET-1 promotes the growth and metastasis of esophageal squamous cell carcinoma via activating PI3K/Akt pathway

    Journal: Translational Cancer Research

    doi: 10.21037/tcr.2020.04.26

    Knockdown of ET-1 blocked the PI3K/Akt pathway. Western blot was performed to detect the level of p-PI3K, PI3K, p-Akt473, p-Akt308 and Akt in ECA109 cells. ***, P<0.001, compared with the control group; ### , P<0.001, compared with the si-ET-1 group.
    Figure Legend Snippet: Knockdown of ET-1 blocked the PI3K/Akt pathway. Western blot was performed to detect the level of p-PI3K, PI3K, p-Akt473, p-Akt308 and Akt in ECA109 cells. ***, P<0.001, compared with the control group; ### , P<0.001, compared with the si-ET-1 group.

    Techniques Used: Western Blot

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    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of <t>p-Akt308</t> and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.
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    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of <t>p-Akt308</t> and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.
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    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of <t>p-Akt308</t> and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.
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    Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of p-Akt308 and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.

    Journal: eLife

    Article Title: ACE2 pathway regulates thermogenesis and energy metabolism

    doi: 10.7554/eLife.72266

    Figure Lengend Snippet: Primary brown adipocytes were isolated, cultured, and treated with Ang-(1-7) (10 –6 M) for 24 hr, Akt inhibitor MK2206 (20 μM) for 24 hr, PKA inhibitor H89 (30 μM) for 24 hr, or adenylylcyclase inhibitor SQ-22536 (10 μM) for 24 hours. ( A, B ) Representative western blots showing the changes of p-Akt308 and Akt in BAT of Ace2 -/y ( A ) and Lepr db/db + Ace2 mice ( B ) exposure at 4 °C (n = 3/each group). ( C ) Representative western blots showing the Akt, UCP1 and forkhead box protein O 1 (FoxO1) Changes (n = 3/each group). ( D ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( E ) Continuous measurement of oxygen consumption rate (OCR) in Ang-(1-7) and MK2206 treated primary brown adipocytes. Oxygen consumption was performed under basal conditions, following the addition of oligomycin (1 μM), the pharmacological uncoupler FCCP (1 μM) or the Complex III and I inhibitor antimycin A and rotenone (0.5 μM) (n = 3–4/each group). ( F, G ) Representative western blots showing the p-PKA and PKA changes in BAT of Ace2 -/y ( F ) and Lepr db/db + Ace2 mice ( G ) exposure at 4 °C (n = 3/each group). ( H ) Relative mRNA levels of thermogenic and mitochondrial genes (n = 4–6/each group). ( I ) Representative western blots showing the UCP1 and PGC1α changes (n = 3/each group). ( J ) Continuous measurement of OCR in Ang-(1-7) and H89-treated primary brown Adipocytes (n = 3–5/each group). Data represent mean ± SEM. *p< 0.05, **p < 0.01 vs GFP/WT group by Student’s t -test. *p < 0.05, **p < 0.01 vs control group, # p < 0.05, ## p < 0.01 vs Ang-(1-7) group by one-way ANOVA. ( K ) Mechanisms involved in ACE2 pathway activation-induced improvement of BAT function.

    Article Snippet: Antibody , Anti-p-Akt308 (rabbit monoclonal) , Cell signaling , #13038 RRID: AB_2629447 , (1:1000).

    Techniques: Isolation, Cell Culture, Western Blot, Activation Assay

    Journal: eLife

    Article Title: ACE2 pathway regulates thermogenesis and energy metabolism

    doi: 10.7554/eLife.72266

    Figure Lengend Snippet:

    Article Snippet: Antibody , Anti-p-Akt308 (rabbit monoclonal) , Cell signaling , #13038 RRID: AB_2629447 , (1:1000).

    Techniques: Transfection, Construct, Sequencing, Software