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Anti phospho s6k thr389

Manufactured by Cell Signaling Technology
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Anti-phospho-S6K (Thr389) is a primary antibody that specifically detects the phosphorylation of S6 kinase (S6K) at threonine 389. S6K is a key downstream effector of the mammalian target of rapamycin (mTOR) signaling pathway, which regulates various cellular processes such as cell growth, proliferation, and metabolism.

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Lab products found in correlation

Anti s6k, by Cell Signaling Technology (6 mentions) Anti akt, by Cell Signaling Technology (4 mentions) Anti mtor, by Cell Signaling Technology (3 mentions) Anti phospho akt ser473, by Cell Signaling Technology (3 mentions) Anti 4e bp1, by Cell Signaling Technology (2 mentions) Anti phospho s6 ser235 236, by Cell Signaling Technology (1 mentions) Anti hif 1α, by BD (1 mentions) Anti muc1, by Cell Signaling Technology (1 mentions) Anti hk2, by Cell Signaling Technology (1 mentions) Anti epas1 hif 2α, by Cell Signaling Technology (1 mentions) Anti rps3, by Cell Signaling Technology (1 mentions) Anti pdk1, by Cell Signaling Technology (1 mentions) α sma, by Merck Group (1 mentions) Anti phospho akt ser473 9271, by Cell Signaling Technology (1 mentions) Anti phospho akt t308, by Cell Signaling Technology (1 mentions) Anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Merck Group (1 mentions) Anti gapdh, by Merck Group (1 mentions) Fibronectin, by Merck Group (1 mentions) Anti fibronectin, by Merck Group (1 mentions)

Close spelling variants of this product

Anti-S6K (42 citations) Phospho-S6K (35 citations) Anti-phospho-S6K (11 citations) Phospho-S6K (Thr389) (4 citations)

6 protocols using anti phospho s6k thr389

1

Immunoblot Analysis of Cellular Signaling

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Immunoblot analysis was conducted as described previously [48 (link)]. Briefly, equal amounts of proteins were resolved on an SDS-polyacrylamide gradient gel and transferred by electroblotting onto a nitrocellulose membrane. Membranes were probed with the indicated primary antibodies. The specific signals were visualized with a chemiluminescence detection system using appropriate secondary antibodies (Perkin-Elmer, Waltham, MA, USA). The following antibodies were used for immunoblotting: anti-TOP2B (Abcam, Cambridge, MA, USA), anti-HIF-1α, anti-ARNT/HIF-1β (BD Transduction, San Jose, CA, USA), anti-EPAS1/HIF-2α, anti-mTOR, anti-MUC1, anti-HK2, anti-PDK1, anti-4EBP1, anti-phospho-4EBP1 (Ser65), anti-S6, anti-phospho-S6 (Ser235/236), anti-S6K, anti-phospho-S6K (Thr389) and anti-RPS3 (Cell Signaling Technology, Danvers, MA, USA). TOP2B, mTOR and RPS3 were used as controls.
Koido M., Haga N., Furuno A., Tsukahara S., Sakurai J., Tani Y., Sato S, & Tomida A. (2017). Mitochondrial deficiency impairs hypoxic induction of HIF-1 transcriptional activity and retards tumor growth. Oncotarget, 8(7), 11841-11854.
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2

Fibrosis Signaling Pathway Protocols

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PDE5-I vardenafil was obtained from Sequoia Research Products Ltd. (vardenafil citrate), Sigma Aldrich, or Selleckchem (vardenafil HCl). Nintedanib was obtained from Cayman Chemical. TGF-β1 was obtained from R&D Systems. The antibodies used in this study were as follows: fibronectin (Sigma Aldrich, F3648, St. Louis, MO, USA), CTGF (Santa Cruz Biotechnology, sc-14939, Dallas, TX, USA), αSMA (Sigma, A2547), p-SMAD3 (Cell Signaling 95205, Danvers, MA, USA), total SMAD3 (Abcam AB 28379), and GAPDH (Millipore MAB374). The secondary antibodies used for fibronectin ELISA included anti-rabbit IgG peroxidase (Sigma Aldrich, A0545) and anti-rabbit IgG-HRP (SantaCruz Biotechnology sc-2004). Additional antibodies for TGF-β1 signaling included the following: anti-mouse serpin E1/PAI-1 antibody (AF3828; R&D systems), anti-CTGF (sc-14939; SCBT), anti-αSMA (A2547; Sigma), anti-fibronectin (F3648, Sigma), anti-GAPDH (AB2302; Millipore, Burlington, MA, USA), anti-phospho-SMAD3 and anti-phospho-SMAD2 antibodies generated in our laboratory, anti-SMAD2 (ab63576; Abcam, Cambridge, UK), anti-SMAD3 (ab28379; Abcam), anti-phospho-AKT (Ser473; 9271, Cell Signaling), anti-phospho-AKT (T308; 4056, Cell Signaling), anti-phospho S6K (Thr389, 9234, Cell Signaling), anti-Akt (9272, Cell Signaling), and anti-S6K (9202, Cell Signaling).
Bourne MH J.r., Kottom T.J., Hebrink D.M., Choudhury M., Leof E.B, & Limper A.H. (2021). Vardenafil Activity in Lung Fibrosis and In Vitro Synergy with Nintedanib. Cells, 10(12), 3502.
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3

Western Blot Antibody Reagents

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Anti-S6K (#9202), anti-phospho S6K Thr389 (#9234), anti-AKT (#9272), anti-phospho AKT Ser473 (#9271), anti-LAT1 (#5347), anti-mTOR (#2983), anti-Raptor (#2280), anti-Flag (#14793), and anti-Rictor (#2140) were purchased from Cell Signaling Technology. Anti-tubulin (05-829) was purchased from Millipore. Anti-β-actin (A700-057) and anti-Flag M2 (F1804) were purchased from Sigma. HA-horseradish peroxidase (HRP) (11-814-150-001) was purchased from Roche. Anti-HA (sc-7392) was purchased from Santa Cruz Biotechnology. Anti-LAMP2 (ab25631) and anti-sodium potassium ATPase (ab76020) were purchased from Abcam.
Tae K., Kim S.J., Cho S.W., Lee H., Cha H.S, & Choi C.Y. (2023). L-Type Amino Acid Transporter 1 (LAT1) Promotes PMA-Induced Cell Migration through mTORC2 Activation at the Lysosome. Cells, 12(20), 2504.
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4

Muscle Fiber Type Analysis

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We obtained ursolic acid (BML-CT105) and tomatidine (BML-GR335) from Enzo Life Sciences. Puromycin (P8833) was obtained from Sigma-Aldrich. [3H]Leucine (120 Ci/mmol) was obtained from American Radiolabeled Chemicals. Anti-myosin heavy chain I (BA-F8), anti-myosin heavy chain IIa (SC-71), anti-myosin heavy chain IIb (BF-F3), and anti-myosin heavy chain IIx (6H1) mouse monoclonal antibodies were obtained from the University of Iowa Developmental Studies Hybridoma Bank. Alexa Fluor 488-conjugated anti-mouse IgG1, Alexa Fluor 350-conjugated anti-mouse IgG2b, and Alexa Fluor 555-conjugated anti-mouse IgM were obtained from Life Technologies. Anti-FLAG mouse monoclonal antibody (F1804) was obtained from Sigma-Aldrich. Anti-puromycin mouse monoclonal antibody (MABE343) was obtained from EMD Millipore. All other antibodies were obtained from Cell Signaling Technology; these included anti-S6K (catalog number 2308S), anti-phospho-S6K (Thr-389) (catalog number 9234S), anti-Akt (catalog number 4691S), anti-phospho-Akt (Ser-473) (catalog number 4060S), anti-eukaryotic translational initiation factor 4E-binding protein 1 (4E-BP1) (catalog number 9644S), anti-phospho-4E-BP1 (Thr-37/46) (catalog number 2855S), and horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (catalog number 7074S) and anti-mouse IgG (catalog number 7076S).
Ebert S.M., Dyle M.C., Bullard S.A., Dierdorff J.M., Murry D.J., Fox D.K., Bongers K.S., Lira V.A., Meyerholz D.K., Talley J.J, & Adams C.M. (2015). Identification and Small Molecule Inhibition of an Activating Transcription Factor 4 (ATF4)-dependent Pathway to Age-related Skeletal Muscle Weakness and Atrophy. The Journal of Biological Chemistry, 290(42), 25497-25511.
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5

Signaling Pathway Antibody Panel Protocol

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Anti-p110α (#4249), anti-phospho-Akt Ser473 (#4060), anti-phospho-Akt Thr308 (#2965), anti-Akt (#4691), anti-phospho-Pras40 Thr246 (#2997), anti-Pras40 (#2691), anti-phospho-GSK3β Ser9 (#9336), anti-GSK3β (#9315), anti-βactin (#4970), anti-phospho-IKKα/β Ser176/180 (#2697), anti-phospho-IκBα Ser32/36 (#9246), anti-IκBα (#9247), anti-phospho NF-Kappa-B p65 Ser536 (#3033), anti-NF-Kappa-B p65 (#8242), anti-AMPKα (#2532), anti-phospho-AMPKα Thr172 (#2535), anti-ACC (#3676), anti-phospho-ACC Ser79 (#3661), anti-S6K (#2708), anti-phospho-S6K Thr389 (#9205), anti-S6 (#2217), anti-phospho-S6 Ser240/244 (#5364), anti-4EBP1 (#9452), anti-phosho-4EBP1 Ser65 (#9451), and anti-TSC2 (#3990) were purchased from Cell Signaling Technologies. Laminin V (#Z0097) and Ki67 (#M7240) were purchased from Dako. Horseradish peroxidase-conjugated anti-rabbit and anti-mouse immunoglobulin antibodies were purchased from Chemicon.
Henry W.S., Laszewski T., Tsang T., Beca F., Beck A.H., McAllister S.S, & Toker A. (2016). Aspirin suppresses growth in PI3K-mutant breast cancer by activating AMPK and inhibiting mTORC1 signaling. Cancer research, 77(3), 790-801.
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6

Western Blot Analysis of Angiogenic Signaling

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HUVEC (2 × 105cells/well) were seeded in 6-well plates and allowed to adhere overnight. Following drug treatment for 24 h, cells were lysed by adding 2× Laemmli buffer containing 4% SDS, 20% glycerol, 10% 2-mercaptoethanol, 0.004% bromophenol blue, 0.125M Tris-HCl, pH 6.8 and the lysates were boiled for 10 min and vortexed. After SDS-PAGE, the proteins were transferred to nitrocellulose membranes (Bio-Rad, Hercules, CA). The blots were blocked with 5% non-fat dried milk at room temperature for 1 h, incubated with the primary antibodies including anti-VEGFR2 (Cell Signaling Technology), anti-phospho-VEGFR2 (Tyr1175, Cell Signaling Technology), anti-FGFR1 (Cell Signaling Technology), anti-mTOR (Cell Signaling Technology), anti-phospho-mTOR (Ser2448, Cell Signaling Technology) anti-S6K (Cell Signaling Technology), anti-phospho-S6K (Thr389, Cell Signaling Technology), anti-PDGFRβ (Santa Cruz Biotechnologies), anti-actin (Santa Cruz Biotechnologies) or anti-α-tubulin (Santa Cruz Biotechnologies) antibodies overnight at 4°C and then incubated with HRP-conjugated secondary antibodies at room temperature for 1 h. The immune-complexes were detected using enhanced chemiluminescence (ECL) detection reagent (GE Healthcare, Pittsburgh, PA).
Shim J.S., Li R.J., Lv J., Head S., Yang E.J, & Liu J.O. (2015). Inhibition of angiogenesis by selective estrogen receptor modulators through blockade of cholesterol trafficking rather than estrogen receptor antagonism. Cancer letters, 362(1), 106-115.
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