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Phospho s6 kinase

Manufactured by Cell Signaling Technology
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Phospho-S6 kinase is a lab equipment product that detects and quantifies the phosphorylation of the S6 ribosomal protein. S6 kinase is a key regulator of cell growth and proliferation. This product can be used to measure the activation status of the S6 kinase signaling pathway.

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Lab products found in correlation

Phospho akt, by Cell Signaling Technology (3 mentions) Bolt bis tris plus polyacrylamide gels, by Thermo Fisher Scientific (2 mentions) Total s6 kinase, by Cell Signaling Technology (2 mentions) Enhanced chemiluminescence, by GE Healthcare (2 mentions) Psd 95, by Thermo Fisher Scientific (2 mentions) Phospho erk, by Cell Signaling Technology (2 mentions) Redd1, by Proteintech (2 mentions) S6 kinase, by Cell Signaling Technology (2 mentions) Phospho mtor xp, by Cell Signaling Technology (2 mentions) Phospho 4ebp1, by Cell Signaling Technology (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions) Bromodeoxyuridine (brdu), by Cell Signaling Technology (1 mentions) Acetyl coa carboxylase, by Cell Signaling Technology (1 mentions) Phospho s6 ribosomal protein s235 236, by Cell Signaling Technology (1 mentions) Phospho erk t202 y204, by Cell Signaling Technology (1 mentions) Total s6 ribosomal protein, by Cell Signaling Technology (1 mentions) Anti cpt1a antibody, by Abcam (1 mentions) Tuberin tsc2, by Cell Signaling Technology (1 mentions) Phospho rsk s380, by Cell Signaling Technology (1 mentions)

6 protocols using phospho s6 kinase

1

Comprehensive Protein Extraction and Analysis

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Total proteins were extracted through 30-minute incubation on ice with Nonidet P-40 lysis buffer containing protease and phosphatase inhibitors, and resolved on Bolt Bis-Tris Plus polyacrylamide gels (Life Technologies). Antibodies against PARP (cat# 9532S), Phospho-Akt (S473; cat# 4060S), Akt (cat# 4685S), Phospho-Erk (T202/Y204; cat# 9101S), Erk1/2 (cat# 9102S), Phospho-S6 ribosomal protein (S235/236; cat# 2211S), total S6 ribosomal protein (cat# 2317S), Phospho-S6 kinase (cat# 9234S), total S6-kinase (cat# 2708S), Tuberin/TSC2 (cat# 4308S), Phospho-RSK (S380) (cat# 9335), total RSK (cat# 9355), Fatty Acid Synthase (cat# 3180S), Acetyl-CoA Carboxylase (cat# 3676S), Stearoyl-CoA desaturase 1 (cat# 2794S), CCTα (cat# 6931S) and BrdU (5292S) were obtained from Cell Signaling Technology (Danvers, MA). Anti-beta actin antibody (cat# A5316) was obtained from Millipore Sigma (St. Louis, MO) and anti-CPT1A antibody (cat# ab128568) from Abcam (Cambridge, MA).
Feng Y., Mischler W.J., Gurung A.C., Kavanagh T.R., Androsov G., Sadow P.M., Herbert Z.T, & Priolo C. (2020). Therapeutic targeting of the secreted lysophospholipase D autotaxin suppresses tuberous sclerosis complex-associated tumorigenesis. Cancer research, 80(13), 2751-2763.
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2

Immunofluorescence Analysis of Autophagy and mTOR Signaling

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The kidney tissue sections were placed in a blocking serum (5% bovine serum albumin in PBS) for 30 min at room temperature. The tissue sections were incubated anti-Atg 5-12 (1:200 dilution; Cell Signaling Technology, Inc., Beverly, MA, USA), phospho-S6 Kinase (1:200 dilution; Cell Signaling Technology), and phospho-p70S6 Kinase (1:200 dilution; Cell Signaling Technology) for 2 h at room temperature. After washing with PBS, the slides were incubated with secondary antibodies (1:200 dilution), conjugated with Alexa Fluor 488 and Alexa Flour 555 (Invitrogen, Waltham, MA, USA). The tissue sections were stained with the nucleic acid stain Hoechst 33342 (DAPI, 1:1000) at room temperature for 2 min. The slides were mounted using a mounting medium (DAKO, Agilent, SantaClara, CA, USA). The stained slides were viewed under a NIKON A1+ confocal microscope (Nikon, Tokyo, Japan). After capturing the image, the quantitative analysis of the expression of Atg 5-12 and the p-S6K area of each group was conducted with the iSolution DT software.
Kim Y.A., Gu H., Gwon M.G., An H.J., Bae S., Leem J., Jung H.J., Park K.K, & Lee S.J. (2022). Synthetic Non-Coding RNA for Suppressing mTOR Translation to Prevent Renal Fibrosis Related to Autophagy in UUO Mouse Model. International Journal of Molecular Sciences, 23(19), 11365.
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3

Immunoblotting for Synaptic Signaling Proteins

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Rodents were rapidly decapitated, and synaptoneurosome-enriched fractions were prepared from prefrontal cortex tissue samples and sonicated in lysis buffer. Protein was electrophoretically separated on a 7.5% SDS PAGE gel and transferred to a PVDF membrane. Blots were incubated in the appropriate primary antibody [phospho-S6 kinase (Thr 389; Cell Signaling 9234; 1:1,000); S6 kinase (Cell Signaling 2708; 1:1,000); phospho-mTOR XP (Ser 2448; Cell Signaling 5536; 1:500); mTOR (Cell Signaling 2983; 1:1,000); phospho-4EBP1 (Thr 37/46; Cell Signaling 2855; 1:500); phospho-ERK (Thr 202/Tyr204; Cell Signaling 4370; 1:1,000); ERK (Cell Signaling 4695; 1:1,000); phospho-Akt (Ser 473; Cell Signaling 4058; 1:1,000); Akt (Cell Signaling 9272; 1:1,000); GAPDH (Advanced Immunochemical 2-RGM2; 1:20,000); REDD1 (Proteintech 10638-1-AP; 1:500); TSC2 (Santa Cruz sc-893; 1:1,000); PSD-95 (Invitrogen 51-6900; 1:1,000] and developed using enhanced chemiluminescence (GE Biosciences). Optical densities of the bands were analyzed using NIH Image J software. For analysis, protein levels were normalized to total protein levels then expressed as a percentage of that in control animals. For figure panels, contrasts have been adjusted linearly for easier viewing of bands.
Ota K.T., Liu R.J., Voleti B., Maldonado-Aviles J.G., Duric V., Iwata M., Dutheil S., Duman C., Boikess S., Lewis D.A., Stockmeier C.A., DiLeone R.J., Rex C., Aghajanian G.K, & Duman R.S. (2014). REDD1 is essential for stress-induced synaptic loss and depressive behavior. Nature medicine, 20(5), 531-535.
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4

Immunoblotting for Synaptic Signaling Proteins

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Rodents were rapidly decapitated, and synaptoneurosome-enriched fractions were prepared from prefrontal cortex tissue samples and sonicated in lysis buffer. Protein was electrophoretically separated on a 7.5% SDS PAGE gel and transferred to a PVDF membrane. Blots were incubated in the appropriate primary antibody [phospho-S6 kinase (Thr 389; Cell Signaling 9234; 1:1,000); S6 kinase (Cell Signaling 2708; 1:1,000); phospho-mTOR XP (Ser 2448; Cell Signaling 5536; 1:500); mTOR (Cell Signaling 2983; 1:1,000); phospho-4EBP1 (Thr 37/46; Cell Signaling 2855; 1:500); phospho-ERK (Thr 202/Tyr204; Cell Signaling 4370; 1:1,000); ERK (Cell Signaling 4695; 1:1,000); phospho-Akt (Ser 473; Cell Signaling 4058; 1:1,000); Akt (Cell Signaling 9272; 1:1,000); GAPDH (Advanced Immunochemical 2-RGM2; 1:20,000); REDD1 (Proteintech 10638-1-AP; 1:500); TSC2 (Santa Cruz sc-893; 1:1,000); PSD-95 (Invitrogen 51-6900; 1:1,000] and developed using enhanced chemiluminescence (GE Biosciences). Optical densities of the bands were analyzed using NIH Image J software. For analysis, protein levels were normalized to total protein levels then expressed as a percentage of that in control animals. For figure panels, contrasts have been adjusted linearly for easier viewing of bands.
Ota K.T., Liu R.J., Voleti B., Maldonado-Aviles J.G., Duric V., Iwata M., Dutheil S., Duman C., Boikess S., Lewis D.A., Stockmeier C.A., DiLeone R.J., Rex C., Aghajanian G.K, & Duman R.S. (2014). REDD1 is essential for stress-induced synaptic loss and depressive behavior. Nature medicine, 20(5), 531-535.
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5

Protein Extraction and Western Blot Protocol

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Total proteins were extracted through 15-minute incubation with Nonidet P-40 lysis buffer and resolved on Bolt Bis-Tris Plus polyacrylamide gels (Life Technologies). Phospho-S6 kinase (cat# 9234S), total S6-kinase (cat# 2708S), and Tuberin (cat# 4308S) antibodies were obtained from Cell Signaling Technology (Danvers, MA). Beta-actin (cat# A5316) was obtained from Sigma (St. Louis, MO). Primary antibodies were diluted 1:1000 in 5% bovine serum albumin or 5% non-fat milk in Tris-buffered saline (TBS) with 0.5% Tween 20, following the manufacturer’s instructions, and probed on nitrocellulose membrane overnight at 4°C.
Verwer E.E., Kavanagh T.R., Mischler W.J., Feng Y., Takahashi K., Wang S., Shoup T.M., Neelamegam R., Yang J., Guehl N.J., Ran C., Massefski W., Cui Y., El-Chemaly S., Sadow P.M., Oldham W.M., Kijewski M.F., El Fakhri G., Normandin M.D, & Priolo C. (2018). [18F]Fluorocholine and [18F]fluoroacetate PET as imaging biomarkers to assess phosphatidylcholine and mitochondrial metabolism in preclinical models of TSC and LAM. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(23), 5925-5938.
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6

Protein Extraction and Analysis

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Cellular proteins were extracted using RIPA lysis buffer (Thermo Fisher Scientific) supplemented with 2 mM NaVO3, 1 mM PMSF, and 5 mM NaF. The plasma and cellular total protein concentrations were quantified by a Pierce BCA Protein Assay kit (Thermo Fisher Scientific). Primary antibodies against IRAK1, AKT, phospho-AKT (Ser473), S6 kinase, phospho-S6 kinase (Ser235/236), and β-actin (1:1000) (Cell Signaling Technology, CST) were used in the analysis. The protein expression level was assessed using an Immobilon Western Chemiluminescent HRP Substrate kit (Millipore, Merck, Taichung, Taiwan) in an ImageQuant LAS4000 Biomolecular Imager (GE Healthcare, Taichung, Taiwan).
Huang Y.N., Chiang S.L., Lin Y.J., Liu S.C., Li Y.H., Liao Y.C., Lee M.R., Su P.H., Tsai F.J., Hung H.C, & Wang C.H. (2021). Long, Noncoding RNA SRA Induces Apoptosis of β-Cells by Promoting the IRAK1/LDHA/Lactate Pathway. International Journal of Molecular Sciences, 22(4), 1720.
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