Anti phospho akt thr308 244f9

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-Phospho-Akt-Thr308 (244F9) is a monoclonal antibody that specifically detects Akt (protein kinase B) phosphorylated at threonine 308. This antibody can be used to detect the activation of the Akt signaling pathway.

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Anti-AKT (1447 citations) Phospho-Akt (892 citations) Anti-phospho-Akt (493 citations) Phospho-AKT (Thr308) (121 citations) Anti-phospho-AKT (Thr308) (53 citations) Phospho-Akt (Thr308) (244F9) (2 citations)

3 protocols using anti phospho akt thr308 244f9

Adipogenic Differentiation of Rat MSCs

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Mesenchymal stromal cells (MSCs) from 12-week old WT and Cyp2j4^−/− rats were obtained as previously described [32] (link). MSCs cells were allowed to grow in Supplemented MesenCult™ MSC Medium (STEMCELL Technologies, UK) for 5 days on Petri dishes (Nunc, ThermoFisher Scientific, UK). MSCs from WT and Cyp2j4^−/− rats were differentiated into mature adipocytes by incubation with an adipogenic induction medium (StemPro®,Gibco, UK) for 14 days.
Antibodies used in western blot were: anti-PPAR-γ (C26 H12, Cell Signaling #2435, 1:1000), anti CEBP-α (Cell Signaling #2295, 1:1000), anti-Phospho-Akt-Ser473 (D9E, Cell Signaling #4060, 1:2000), anti-Phospho-Akt-Thr308 (244F9, Cell Signaling #4056, 1:1000), anti-panAkt (C67E7, Cell Signaling #4691, 1:1000) and anti-β-Actin Antibody (C4, sc-47778, 1:10,000), anti-PPARα (H2, SC-398,394, 1:1000), anti-PPARβ/δ (F-10, SC-74517, 1:1000), anti-FXR (D-3, SC-25309, 1:1000), anti-LXRα (ab2585, 1:1000), and anti-β-Actin Antibody (C4, sc-47778, 1:10,000).

Olona A., Terra X., Ko J.H., Grau-Bové C., Pinent M., Ardevol A., Diaz A.G., Moreno-Moral A., Edin M., Bishop-Bailey D., Zeldin D.C., Aitman T.J., Petretto E., Blay M, & Behmoaras J. (2018). Epoxygenase inactivation exacerbates diet and aging-associated metabolic dysfunction resulting from impaired adipogenesis. Molecular Metabolism, 11, 18-32.

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Targeted Cancer Therapy Compounds and Antibodies

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The following drugs were purchased: Dinaciclib (SCH727965) for in vitro and in vivo studies (S2768; Selleckchem), lapatinib ditosylate (Tykerb) for in vitro and in vivo studies (M1802; Abmole), neratinib for in vivo studies (M1913; Abmole), A-1210477 (CT-A121; Chemietek), A-1331852 (22963; Cayman Chemicals), tucatinib (HY-16069; Medchem), and ABT-199 (venetoclax) (CT-A199; Chemietek). The antibodies used in this study were as follows: anti-Bak (AB-1 clone for IP) (AM03; EMD Millipore), anti-Bak (3814S; Cell Signaling), anti-Bim (C34C5) (2933S; Cell Signaling), anti–BCL-xL (54H6) (2764S; Cell Signaling), anti–Bcl-2 (D55G8) (Human Specific) (4223S; Cell Signaling), anti-cleaved PARP (Asp214) (D64E10) (5625S; Cell Signaling), anti-GAPDH (6C5) (sc-32233; Santa Cruz), anti–MCL-1 (S-19) (sc-819; Santa Cruz), anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) (4370S; Cell Signaling), anti-phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) (5364S; Cell Signaling), anti-phospho-Akt (Thr308) (244F9) (4056S; Cell Signaling), anti-phospho-Akt (Ser473) (D9E) (4060S; Cell Signaling), anti-HER2/ErbB2 (29D8) (2165S; Cell Signaling), anti-phospho-HER2/ErbB2 (Tyr1248) (2247S; Cell Signaling), anti-phospho-Rpb1 CTD (Ser 2/5) (4375S; Cell Signaling), Normal Rabbit IgG for IP (sc-2027; Santa Cruz), and Normal Mouse IgG for IP (sc-2025; Santa Cruz).

Floros K.V., Jacob S., Kurupi R., Fairchild C.K., Hu B., Puchalapalli M., E. Koblinski J., Dozmorov M.G., Boikos S.A., Scaltriti M, & Faber A.C. (2021). Targeting transcription of MCL-1 sensitizes HER2-amplified breast cancers to HER2 inhibitors. Cell Death & Disease, 12(2), 179.

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Quantification of Akt and S6 Kinase Activation

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Membranes were incubated with anti-phospho-Akt (Thr308)(244F9) (#4056, Cell Signaling Technology, Danvers, MA, USA), anti-phospho-Akt (Ser473) (#9271, Cell Signaling Technology, Danvers, MA, USA), Akt1/2 Antibody (N-19) (#Sc-1619, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Phospho-p70 S6 Kinase (Thr389)(108D2) (#9234, Cell Signaling Technology, Danvers, MA, USA), p70 S6 Kinase (#9202, Cell Signaling Technology, Danvers, MA, USA), phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) (#5364, Cell Signaling Technology, Danvers, MA, USA), S6 Ribosomal Protein (5G10) (#2217, Cell Signaling Technology, Danvers, MA, USA), anti-P-glycoprotein C219 (#517310, Merck Millipore, Burlington, MA, USA), anti-BCRP/ABCG2 antibody BXP-21 (#ab3380, Abcam, Cambridge, MA, USA), or anti-α-tubulin (#T6199, Sigma-Aldrich, St. Louis, MO, USA) primary antibody and subsequently incubated with horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G (IgG) or anti-rabbit IgG secondary antibodies. Signals were detected as described previously [44 (link)].

Wu C.P., Hung C.Y., Lusvarghi S., Huang Y.H., Tseng P.J., Hung T.H., Yu J.S, & Ambudkar S.V. (2020). Overexpression of ABCB1 and ABCG2 contributes to reduced efficacy of the PI3K/mTOR inhibitor samotolisib (LY3023414) in cancer cell lines. Biochemical pharmacology, 180, 114137.

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