P70 s6 kinase thr389

Manufactured by Cell Signaling Technology

Sourced in United States

P70 S6 Kinase (Thr389) is a lab equipment product that serves as a phosphorylation site-specific antibody. It is used to detect and analyze the phosphorylation of the Thr389 residue on the P70 S6 kinase protein, which is an important regulator of cell growth and proliferation.

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Lab products found in correlation

Supersignal west femto maximum sensitivity substrate kit, by Thermo Fisher Scientific (2 mentions) Molecular imager chemidoc xrs system, by Bio-Rad (2 mentions) Eif4g ser1108, by Cell Signaling Technology (2 mentions) Rvfv mp12 antibody, by IBT Bioservices (2 mentions) Rapamycin, by Merck Group (1 mentions) Phalloidin 488, by Thermo Fisher Scientific (1 mentions) P erk1 2 t202 y204, by Cell Signaling Technology (1 mentions) Cambinol, by Merck Group (1 mentions) Mitotracker red, by Thermo Fisher Scientific (1 mentions) Ex527, by Selleck Chemicals (1 mentions) Lc3 1 2, by Cell Signaling Technology (1 mentions) Actinomycin d, by Merck Group (1 mentions) Nicotinamide, by Merck Group (1 mentions) Gm130, by BD (1 mentions) Lamp1 antibody h4a3, by Developmental Studies Hybridoma Bank (1 mentions) P90rsk ser380, by Cell Signaling Technology (1 mentions) Ab49900, by Abcam (1 mentions) Caov3, by American Type Culture Collection (1 mentions) P p70 s6 kinase thr389, by Cell Signaling Technology (1 mentions) Skov3, by American Type Culture Collection (1 mentions)

5 protocols using p70 s6 kinase thr389

Multimodal Analysis of Autophagy Regulation

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Cambinol, Actinomycin D, TSA, Rapamycin, and Nicotinamide were purchased from Sigma (St. Louis, MO). AK1 and AGK2 were purchased from ChemBridge (San Diego, Ca). EX527 was supplied by Selleck Chemicals (Houston, TX). UO126 was purchased from Calbiochem (Billerica, MA) and used at 10 μM. LAMP-1 antibody (h4A3) was obtained from the Developmental Studies Hybridoma Bank at the University of Iowa and used at a 1:200 dilution. GM130 and EEA1 antibodies (BD Biosciences, San Jose, CA) were used at 1:50. Mitotracker Red (Molecular Probes, Carlsbad, CA) was used according to the manufacturer's protocol. The SIRT1 (B-10) and Ac-tubulin antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX) and was used at 1:1000. pMEK1/2 S271/221, p70 S6 Kinase Thr389, pErk1/2 T202/Y204, Ac-p53 Lys382, LC3I/II, and total Erk1/2 antibodies (Cell Signaling Technology, Beverly, MA) were used at 1:1000 for western blot. Anti-tubulin antibody (NeoMarkers, Fremont, CA) was used at 1:100 for immunofluorescence (IF) and 1:20,000 for western blot analysis. Secondary antibodies include Dylight 594 donkey anti-mouse 1:100 (Jackson IR, West Grove, PA) for immunoflorescence, and HRP-conjugated anti-mouse and anti-rabbit for western blot (GE Healthcare, Pittsburgh, PA) used at 1:5000. Phalloidin 488 (Invitrogen, Carlsbad, CA) was used at 1:200.

Dykes S.S., Friday E., Pruitt K, & Cardelli J.A. (2015). The histone deacetylase inhibitor cambinol prevents acidic pHe-induced anterograde lysosome trafficking independently of sirtuin activity. Biochemistry and Biophysics Reports, 3, 83-93.

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Protein Expression Analysis by Western Blot

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Protein lysates were collected and analyzed by western blot as previously described [42 (link)]. In brief, primary antibodies against p90RSK (Ser380) (Cell Signaling Technology, Danvers, MA, USA, 9341), p70 S6 Kinase (Thr389) (Cell Signaling Technology 9205), S6 ribosomal protein (Ser235/236) (Cell Signaling Technology, 4856), eIF4G (Ser1108) (Cell Signaling Technology, 2441), RVFV MP12 Antibody (IBT Bioservices, Rockville, MD, USA, 04-0001), or HRP-conjugated actin (Abcam, ab49900) were diluted 1:1000 in 5% bovine serum albumin in 1× TBS with 0.1% Tween-20 solution followed by the addition of the appropriate secondary antibody. The western blots were visualized by chemiluminescence using SuperSignal West Femto Maximum Sensitivity Substrate kit (ThermoScientific, Waltham, MA, USA) and a Bio Rad Molecular Imager ChemiDoc XRS system (Bio-Rad, Hercules, CA, USA) [41 (link),42 (link)].

Bell T.M., Espina V., Lundberg L., Pinkham C., Brahms A., Carey B.D., Lin S.C., Dahal B., Woodson C., de la Fuente C., Liotta L.A., Bailey C.L, & Kehn-Hall K. (2018). Combination Kinase Inhibitor Treatment Suppresses Rift Valley Fever Virus Replication. Viruses, 10(4), 191.

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Western Blot Analysis of Signaling Proteins

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Protein lysates were collected and analyzed by western blot as previously described (Austin et al., 2012 (link)). In brief, primary antibodies against p70 S6 Kinase (Thr389) (Cell Signaling Technology 9205), S6 ribosomal protein (Ser235/236) (Cell Signaling Technology, 4856), eIF4G (Ser1108) (Cell Signaling Technology, 2441), RVFV MP12 Antibody (IBT Bioservices 04-0001), or HRP-conjugated actin (Abcam, ab49900) were diluted 1:1000 in 5% bovine serum albumin in 1× TBS with 0.1% Tween-20 solution followed by the addition of the appropriate secondary antibody. The western blots were visualized by chemiluminescence using SuperSignal West Femto Maximum Sensitivity Substrate kit (ThermoScientific) and a Bio Rad Molecular Imager ChemiDoc XRS system (Bio-Rad) (Austin et al., 2012 (link); Baer et al., 2012 (link)).

Bell T.M., Espina V., Senina S., Woodson C., Brahms A., Carey B., Lin S.C., Lundberg L., Pinkham C., Baer A., Mueller C., Chlipala E.A., Sharman F., de la Fuente C., Liotta L, & Kehn-Hall K. (2017). Rapamycin modulation of p70 S6 kinase signaling inhibits Rift Valley fever virus pathogenesis. Antiviral research, 143, 162-175.

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Resistin Regulates Ovarian Cancer Progression

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Two human ovarian epithelial carcinoma cell lines (CAOV3 and SKOV3) were purchased from ATCC (Manassas, VA, USA). McCoy5A medium was purchased from HyClone (Logan, UT, USA). Recombinant resistin, mTOR antibody, rapamycin, adult fat mesenchymal stem cells, and adult fat mesenchymal stem cells complete medium were purchased from Phoenix Pharmaceuticals (Phoenix Pharmaceuticals, Mannheim, Germany). Anti-βactin antibody was obtained from Keygen Biotech (1:1000, Nanjing, China). Phospho (p)- mTOR, mTOR, p- p70S6 kinase (Thr389), and p70S6 kinase (Thr389) were purchased from Cell Signaling Technology (1:1000, Danvers, MA, USA). Cell Counting Kit-8 (CCK-8) was supplied by Dojindo (Kumamoto, Japan), and the ELISA plate reader was obtained from Bio-Rad (Hercules, CA, USA).

Pang L, & Chang X. (2021). Resistin Expression in Epithelial Ovarian Cancer promotes the Proliferation and Migration of Ovarian Cancer Cells to Worsen Prognosis. Journal of Cancer, 12(22), 6796-6804.

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Tumor Protein Extraction and Analysis

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Snap-frozen tumor tissues were homogenized in RIPA buffer (20 mM Tris–HCl pH 8.0, 150 mM NaCl, 1 mM disodium EDTA, 1 mM EGTA, 2.5 mM sodium pyrophosphate, 1 mM β-glycerophosphate, 1% Triton-X100) plus 1 mM phenylmethylsulphonyl fluoride and Halt protease and phosphatase inhibitor cocktail (Thermo Scientific), and handled as we described for the bullet blender protocol (20 (link)) (5 min at speed 10 with beads; Next Advance). Lysates were centrifuged at 14,000 rpm, 4° C for 10 min, transferred into pre-chilled Eppendorf tubes and protein concentrations were determined by the Bradford method (Thermo). 50 μg protein was separated on 4–15% sodium dodecyl sulfate polyacrylamide gels (BioRad), transferred to PVDF membranes (GE Technologies), blocked in Tris-buffered saline (pH 7.4) plus 0.1% Tween-20 and 5% skim milk, and incubated overnight at 4° C with 1:1000 diluted phospho- and/or total antibodies against rpS6^S240/244, 4E-BP1^T37/46, P70 S6 Kinase^{Thr 389}, and Akt^S473 (all from Cell Signaling) plus anti-mouse β-Actin or α-Tubulin (Santa Cruz Biotechnology) as a loading control. Membranes were incubated with HRP-conjugated antibodies at ambient temperature for 1 h. Proteins were detected by enhanced chemiluminescence (Pierce). Band quantification was done with Image J software (National Institutes of Health).

Liu Y., Pandeswara S., Dao V., Padrón Á., Drerup J.M., Lao S., Liu A., Hurez V, & Curiel T.J. (2016). Biphasic rapamycin effects in lymphoma and carcinoma treatment. Cancer research, 77(2), 520-531.

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