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Pakt2 s474

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PAKT2 S474 is a laboratory reagent that detects the phosphorylation of the AKT2 protein at serine 474. It is a tool for researchers to study cellular signaling pathways involving the AKT2 protein.

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Lab products found in correlation

Pakt1 s473, by Cell Signaling Technology (2 mentions) Pakt s473, by Cell Signaling Technology (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions) Pan akt, by Cell Signaling Technology (1 mentions) Ps6 s240 244, by Cell Signaling Technology (1 mentions) Pakt t308, by Cell Signaling Technology (1 mentions) Recombinant human egf, by Thermo Fisher Scientific (1 mentions) Rad001, by Selleck Chemicals (1 mentions) Antibody against hsc 70, by Santa Cruz Biotechnology (1 mentions) Pgsk3α β s21 9, by Cell Signaling Technology (1 mentions) Pher2 y877, by Cell Signaling Technology (1 mentions) Mmp 2, by Cell Signaling Technology (1 mentions) Hrp conjugated igg secondary antibody, by Cell Signaling Technology (1 mentions) Image lab software, by Bio-Rad (1 mentions) Pvdf membrane, by GE Healthcare (1 mentions) Ecl plus western blotting detection system, by GE Healthcare (1 mentions) β actin, by Merck Group (1 mentions) Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions) Total akt, by Cell Signaling Technology (1 mentions)

5 protocols using pakt2 s474

1

Comprehensive Signaling Pathway Analysis

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Antibodies against AKT1, AKT2, AKT3, panAKT, pAKT S473, pAKT T308, pAKT1 S473, pAKT2 S474, pS6 S240/244, pGSK3α/β S21/9, pHER2 Y877, S100A4, MMP2, RANK, and CTGF, and secondary HRP-linked antibodies against rabbit or mouse IgG were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Antibody against AKT3 was provided by Millipore (Burlington, MA, USA). Antibody against HSC-70 was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Antibody against pDDR1/2 Y796/Y740 was provided by R&D systems Inc. (Minneapolis, MN, USA). RAD001 was provided by Selleck Chemicals (Houston, TX, USA) and MK-2206 was obtained from AbMole BioScience Inc. (Houston, TX, USA). Recombinant human TGF-β1 and recombinant human EGF were purchased from PeproTech Inc. (Rocky Hill, NJ, USA).
Hinz N., Baranowsky A., Horn M., Kriegs M., Sibbertsen F., Smit D.J., Clezardin P., Lange T., Schinke T, & Jücker M. (2021). Knockdown of AKT3 Activates HER2 and DDR Kinases in Bone-Seeking Breast Cancer Cells, Promotes Metastasis In Vivo and Attenuates the TGFβ/CTGF Axis. Cells, 10(2), 430.
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2

Quantitative Western Blot Analysis

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Protein concentration for the resulting lysates was determined using the BCA (bicinchoninic acid) Protein Assay kit from Pierce, and lysates were run on 10% polyacrylamide gels. PVDF membranes (GE healthcare) were incubated with following antibodies: HO-1 (Abcam), β-actin (Sigma), pAkt2-S474 and Akt2 (all Cell Signaling). Hrp-conjugated IgG secondary antibodies were used (Cell Signaling) and blots were developed with ECL Plus Western Blotting Detection System (GE Healthcare). ImageLab software (BioRad) was used for densitometric quantification.
Wagner G., Lindroos-Christensen J., Einwallner E., Husa J., Zapf T.C., Lipp K., Rauscher S., Gröger M., Spittler A., Loewe R., Gruber F., Duvigneau J.C., Mohr T., Sutterlüty-Fall H., Klinglmüller F., Prager G., Huppertz B., Yun J., Wagner O., Esterbauer H, & Bilban M. (2017). HO-1 inhibits preadipocyte proliferation and differentiation at the onset of obesity via ROS dependent activation of Akt2. Scientific Reports, 7, 40881.
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3

Investigating AKT Signaling in Human Skeletal Myoblasts

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Human Skeletal Myoblasts (#A11440 or #A12555), low glucose Dulbecco's Modified Eagle Medium (#11885092), Horse Serum (#26050070), Bovine Serum Albumin (#A9576), and Pierce A/G Magnetic Beads (#88802) were purchased from Thermofisher Scientific Antibodies for Total AKT1 (#2938), Total AKT2 (#2964), Total AKT3 (#14293), Total AKT (#4685), p‐AKT S473 (#4060), p‐AKT1 S473 (#9018), p‐AKT2 S474 (#8599), AKT (Pan) (C67E7) (rabbit monoclonal; #4691), AKT (Pan) (40D4) (mouse monoclonal; #2920), and Human Insulin‐like Growth Factor 1 (#8917) were purchased from Cell Signaling Technologies. Secondary antibodies Anti‐Rabbit IgG HRP‐Linked Antibody (PN#7074) and Anti‐Mouse IgG HRP‐Linked Antibody (PN#7076) were purchased from Cell Signaling Technologies. Human Insulin Solution (#I9278) was purchased from Sigma Aldrich ImageJ was downloaded from the National Institutes of Health website.
Roberts B.M., Geddis A.V, & Matheny RW J.r. (2023). Differential activation of AKT isoforms by growth factors in human myotubes. Physiological Reports, 11(20), e15805.
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4

Evaluation of mTOR and PI3K/Akt Signaling

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Antibodies against phospho-mTOR (S2448, Cat# 2971), mTOR, phospho-S6 (S235/236, Cat# 4856), S6, phospho-Akt1 (S473, Cat# 9271), phospho-Akt1 (T308, Cat# 9275), phospho-GSK-3α/β (S21/9, Cat# 8566), GSK-3α/β (Cat# 5676), phospho-PRAS40 (T246, Cat# 2997), PRAS40 (Cat# 2691), P-Akt2 (S474, Cat# 8599), Akt2 (Cat# 5239) and GAPDH (Cat# 2118) were purchased from Cell Signaling Technology (Frankfurt, Germany). Rapamycin was obtained from Enzo Life Science (Loerrach, Germany). The PI3K inhibitor LY294002 was purchased from Calbiochem (Schwalbach, Germany). The Akt inhibitor MK2206 was purchased from Selleck Chemicals (Munich, Germany). SMART pool siRNA against AKT1 (Cat. No. M-0030000-03) and AKT2 (Cat. No. M-003001-02), as well as a non-targeting siRNA (Cat. No. D-001810-10) were purchased from Thermo Scientific Dharmacon (Bonn, Germany). Anti-Akt1 antibody, anti-phospho-histone H2AX (S139) antibody and the Alexa Fluor 488 conjugated secondary antibody have been previously described [16 (link), 23 (link)].
Holler M., Grottke A., Mueck K., Manes J., Jücker M., Rodemann H.P, & Toulany M. (2016). Dual Targeting of Akt and mTORC1 Impairs Repair of DNA Double-Strand Breaks and Increases Radiation Sensitivity of Human Tumor Cells. PLoS ONE, 11(5), e0154745.
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5

LPS-induced CREB and NF-κB signaling

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BMDMs (3 × 106 per well in 6-well cell culture plates) or RAW264.7 cells (2 × 106 per well in 6-well cell culture plates) were treated with LPS 10 or 100 ng/ml for 30 min and lysed in RIPA buffer with protease and phosphatase inhibitors (Roche, Nutley, NJ). Western blot analysis was performed with primary antibodies against p-CREB-S133, CREB, IRS1, IRS2, p-AKT-S473, AKT, p-AKT1-S473, AKT1, p-AKT2-S474, AKT2, p-NF-κB P65-S536, NF-κB P65, IκBα, β-actin, and GAPDH from Cell Signaling Technology (Denvers, MA) and GHSR from Invitrogen (Waltham, MA). Signaling was visualized with ECL (Genesee Scientific, San Diego, CA) and analyzed using Image J software.
Kim D.M., Lee J.H., Pan Q., Han H.W., Shen Z., Eshghjoo S., Wu C.S., Yang W., Noh J.Y., Threadgill D.W., Guo S., Wright G., Alaniz R, & Sun Y. (2023). Nutrient-sensing growth hormone secretagogue receptor in macrophage programming and meta-inflammation. Molecular Metabolism, 79, 101852.
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