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rabbit polyclonal anti p akt 473  (Proteintech)


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    Proteintech rabbit polyclonal anti p akt 473
    Rabbit Polyclonal Anti P Akt 473, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 527 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rabbit+polyclonal+anti+p+akt+473/Phospho-AKT1+(Ser473)+Antibody/pmc11727224-91-35-42
    Average 96 stars, based on 527 article reviews
    rabbit polyclonal anti p akt 473 - by Bioz Stars, 2026-10
    96/100 stars

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    Membrane:

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis.
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene fluoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% nonfat milk for 1 h at room temperature, and subsequently incubated overnight at 4 °C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-β-Actin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRP-conjugated secondary antibody at room temperature for 1 h, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene fluoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% non-fat milk for 1 h at room temperature, and subsequently incubated overnight at 4 °C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-β-Actin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRP-conjugated secondary antibody at room temperature for 1 h, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene uoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% nonfat milk for 1 h at room temperature, and subsequently incubated overnight at 4°C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-βActin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRPconjugated secondary antibody at room temperature for 1 hour, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.

    Saline:

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis.
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene fluoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% nonfat milk for 1 h at room temperature, and subsequently incubated overnight at 4 °C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-β-Actin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRP-conjugated secondary antibody at room temperature for 1 h, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene fluoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% non-fat milk for 1 h at room temperature, and subsequently incubated overnight at 4 °C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-β-Actin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRP-conjugated secondary antibody at room temperature for 1 h, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene uoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% nonfat milk for 1 h at room temperature, and subsequently incubated overnight at 4°C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-βActin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRPconjugated secondary antibody at room temperature for 1 hour, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.

    Incubation:

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis.
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene fluoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% nonfat milk for 1 h at room temperature, and subsequently incubated overnight at 4 °C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-β-Actin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRP-conjugated secondary antibody at room temperature for 1 h, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene fluoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% non-fat milk for 1 h at room temperature, and subsequently incubated overnight at 4 °C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-β-Actin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRP-conjugated secondary antibody at room temperature for 1 h, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.

    Article Title: TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis
    Article Snippet: Equal amounts of protein (20 μg) from each sample were added to 4× Laemmli sample buffer and subjected to sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) (Invitrogen), followed by transfer onto a polyvinylidene uoride (PVDF) membrane (Millipore, Boston, MA, USA). .. The membrane was washed with TBST buffer (Tris-buffered saline containing 0.1% Tween 20), blocked with 5% nonfat milk for 1 h at room temperature, and subsequently incubated overnight at 4°C with primary antibodies, including rabbit polyclonal anti-p-Akt 473 (1:1000, Cat# 28731-1-AP, Proteintech), rabbit polyclonal anti-Akt (1:1000, Cat# 10176-2-AP, Proteintech), rabbit monoclonal anti-βActin (1:5000, Cat# ab213262, Abcam), mouse monoclonal anti-mTOR (1:1000, Cat# 66888-1-Ig, Proteintech), mouse monoclonal anti-p-mTOR (Ser2448) (1:1000, Cat# 67778-1-Ig, Proteintech), and rabbit polyclonal anti-Hif-1α (1:1000, Cat# 20960-1-AP, Proteintech) antibodies. .. Following primary antibody incubation, the membrane was subjected to additional washes, incubated with an HRPconjugated secondary antibody at room temperature for 1 hour, and then subjected to further washing before visualization via enhanced chemiluminescence using the Bio-Rad ChemiDoc MP imaging system.



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    Image Search Results


    miR-9-5p/Klf4 axis enhances HCC cell proliferation via an AKT/mTOR-associated pathway. (A) The transfection efficiency of miR-9-5p mimic or inhibitor, or siKlf4/Klf4 was measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis. (B) Cell proliferation viability was determined by cell counting kit-8 assay. A total number of 2,000 transfected cells dissolved in 100 µ l 10% fetal bovine serum were seeded into 96-well plates on days 1-5, and the absorbance was measured at 450 nm every 24 h to obtain a cell growth curve. (C) Flow cytometric analysis of SK-Hep-1 and HCC-LM3 cells treated with miR-9-5p mimic or inhibitor, or siKlf4/Klf4 for 48 h. Representative data are featured, presenting the population of living cells (Annexin V-FITC-/PI-) in the left lower quadrant, early apoptotic cells (Annexin V-FITC+/PI-) in right lower quadrant, late apoptotic cells (Annexin V-FITC +/PI+) in the right upper quadrant and necrotic cells (Annexin V-FITC -/PI+) in the left upper quadrant. (D) Levels of AKT, p-AKT S473, mTOR, p-mTOR, Bcl-2 and Bax were examined by western blotting in SK-Hep-1 and HCC-LM3 cells transfected with miR-9-5p mimic/inhibitor or Klf4/siKlf4. (E) Flow cytometric analysis of SK-Hep-1 and HCC-LM3 cells treated with miR-9-5p mimic/inhibitor together with AKTi (10 µ l, MK-2206, 2 HCl; dissolved in DMSO) for 48 h. (F) Cell proliferation analysis of SK-Hep-1 and HCC-LM3 cells treated with miR-9-5p mimic/inhibitor with AKT inhibitor (10 µ l, MK-2206, 2 HCl; dissolved in DMSO) for 48 h. * P<0.05; ** P<0.01; *** P<0.001. miR, microRNA; p, phos-phorylated; AKT, protein kinase B; mTOR, mechanistic target of rapamycin; DMSO, dimethyl sulfoxide; HCC, hepatocellular carcinoma; Klf4, Krüppel-like factor 4; siKlf4, small interfering RNA against Klf4; PI, propidium iodide; FITC, fluorescein isothiocyanate; Bcl-2, B cell lymphoma-2; Bax, Bcl-2-associated X protein; OD, optical density; AKTi, AKT inhibitor.

    Journal: International Journal of Molecular Medicine

    Article Title: MicroRNA-9-5p downregulates Klf4 and influences the progression of hepatocellular carcinoma via the AKT signaling pathway

    doi: 10.3892/ijmm.2019.4062

    Figure Lengend Snippet: miR-9-5p/Klf4 axis enhances HCC cell proliferation via an AKT/mTOR-associated pathway. (A) The transfection efficiency of miR-9-5p mimic or inhibitor, or siKlf4/Klf4 was measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis. (B) Cell proliferation viability was determined by cell counting kit-8 assay. A total number of 2,000 transfected cells dissolved in 100 µ l 10% fetal bovine serum were seeded into 96-well plates on days 1-5, and the absorbance was measured at 450 nm every 24 h to obtain a cell growth curve. (C) Flow cytometric analysis of SK-Hep-1 and HCC-LM3 cells treated with miR-9-5p mimic or inhibitor, or siKlf4/Klf4 for 48 h. Representative data are featured, presenting the population of living cells (Annexin V-FITC-/PI-) in the left lower quadrant, early apoptotic cells (Annexin V-FITC+/PI-) in right lower quadrant, late apoptotic cells (Annexin V-FITC +/PI+) in the right upper quadrant and necrotic cells (Annexin V-FITC -/PI+) in the left upper quadrant. (D) Levels of AKT, p-AKT S473, mTOR, p-mTOR, Bcl-2 and Bax were examined by western blotting in SK-Hep-1 and HCC-LM3 cells transfected with miR-9-5p mimic/inhibitor or Klf4/siKlf4. (E) Flow cytometric analysis of SK-Hep-1 and HCC-LM3 cells treated with miR-9-5p mimic/inhibitor together with AKTi (10 µ l, MK-2206, 2 HCl; dissolved in DMSO) for 48 h. (F) Cell proliferation analysis of SK-Hep-1 and HCC-LM3 cells treated with miR-9-5p mimic/inhibitor with AKT inhibitor (10 µ l, MK-2206, 2 HCl; dissolved in DMSO) for 48 h. * P<0.05; ** P<0.01; *** P<0.001. miR, microRNA; p, phos-phorylated; AKT, protein kinase B; mTOR, mechanistic target of rapamycin; DMSO, dimethyl sulfoxide; HCC, hepatocellular carcinoma; Klf4, Krüppel-like factor 4; siKlf4, small interfering RNA against Klf4; PI, propidium iodide; FITC, fluorescein isothiocyanate; Bcl-2, B cell lymphoma-2; Bax, Bcl-2-associated X protein; OD, optical density; AKTi, AKT inhibitor.

    Article Snippet: The membranes were then treated with 5% non-fat milk blocking solution for 2 h at room temperature, and incubated overnight at 4°C with the following primary antibodies (all Cell Signaling Technology, Inc., Danvers, MA, USA): Anti-Klf4 (cat. no. 4038; 54 kDa; rabbit polyclonal; 1:1,000), anti-AKT Ser-473 (cat. no. 4091; 56 kDa; rabbit polyclonal; 1:1,000), antibody phosphorylated (p)-AKT Ser-473 (cat. no. 4069; 56 kDa; rabbit polyclonal; 1:1,000), anti-Bcl-2 (cat. no. 15071; 26 kDa; mouse polyclonal; 1:1,000), anti-Bax (cat. no. 5032; 20 kDa; rabbit polyclonal; 1:1,000), anti-mTOR (cat. no. 2972; 289 kDa; rabbit poly-clonal; 1:1,000), antibody p-mTOR (cat. no. 5536; 289 kDa; rabbit polyclonal; 1:1,000) and GAPDH (cat. no. 5174; 37 kDa; rabbit monoclonal; 1:5,000).

    Techniques: Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Cell Counting, Small Interfering RNA