Torin

Manufactured by Bio-Techne

Sourced in United Kingdom

Torin is a compact and versatile laboratory centrifuge designed for a wide range of applications. It is capable of handling sample volumes up to 15 mL at speeds up to 6,000 rpm. The Torin centrifuge features a durable metal housing and easy-to-use controls for efficient and reliable operation.

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Lab products found in correlation

Actinomycin d, by Merck Group (3 mentions) Pd0325901, by Merck Group (2 mentions) Sb2035800, by Selleck Chemicals (2 mentions) Cycloheximide (chx), by Merck Group (2 mentions) Anti p70 s6 kinase, by Cell Signaling Technology (1 mentions) Anti mtor antibody, by Cell Signaling Technology (1 mentions) Anti tsc2, by Cell Signaling Technology (1 mentions) Anti pten, by Cell Signaling Technology (1 mentions) Lysotracker, by Thermo Fisher Scientific (1 mentions) Rapamycin, by Merck Group (1 mentions) Wortmannin, by Merck Group (1 mentions) Bafilomycin, by Merck Group (1 mentions) Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions) Oligomycin, by Merck Group (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Tnf α, by Merck Group (1 mentions) Antimycin a, by Merck Group (1 mentions) Doxorubicin, by Merck Group (1 mentions) Uc2288, by Abcam (1 mentions) Arpe 19, by American Type Culture Collection (1 mentions)

12 protocols using torin

Molecular Pathway Regulation in Cancer

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Anti-phopsho-p70S6-kinase, anti-p70S6-kinase, anti-TSC2, anti-PTEN, and anti-mTOR antibodies were purchased from Cell Signaling (Beverly, MA). Anti-LC3 and anti-actin monoclonal antibodies were purchased from Sigma (St. Louis, MO). The heparanase inhibitor PG545 was kindly provided by Progen Pharmaceuticals (Brisbane, Australia) (26 (link)). Cisplatin and doxorubicin were obtained from the Oncology Department, Rambam Health Care Campus (Haifa, Israel). LysoTracker was purchased from Molecular Probes (Life Technologies, Grand Island, NY); Torin was purchased from Tocris Bioscience (Bristol, UK).

Shteingauz A., Boyango I., Naroditsky I., Hammond E., Gruber M., Doweck I., Ilan N, & Vlodavsky I. (2015). Heparanase enhances tumor growth and chemo-resistance by promoting autophagy. Cancer research, 75(18), 3946-3957.

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Autophagy Modulation in Huntington's Disease

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Six Ctrl (1, 2, 4, 6, 8, 10) and six Huntington's disease (1, 2, 5, 6, 9, 10) cell lines, which only included those Huntington's disease lines with the shorter CAG-repeats, were treated with factors regulating autophagy as follows. The cell medium was aspirated from the wells and fresh medium with one of the factors (Bafilomycin, 200 nM, Merck Millipore; Rapamycin, 20 nM, Sigma-Aldrich; Wortmannin, 100 nM, Sigma-Aldrich) was added to the well followed by fixation for immunocytochemistry after 4 h. Torin (250 nM, Tocris Bioscience) treatment was performed identically and only lasted for 2 h. Non-treated wells received fresh media with dimethyl sulphoxide (DMSO) in equivalent amount to that used in treated cells.
Cells were starved by replacing the media with Hank's Balanced Salt Solution (Thermo Fisher, 14025092) for 2 h before fixation.

Pircs K., Drouin-Ouellet J., Horváth V., Gil J., Rezeli M., Garza R., Grassi D.A., Sharma Y., St-Amour I., Harris K., Jönsson M.E., Johansson P.A., Vuono R., Fazal S.V., Stoker T., Hersbach B.A., Sharma K., Lagerwall J., Lagerström S., Storm P., Hébert S.S., Marko-Varga G., Parmar M., Barker R.A, & Jakobsson J. (2021). Distinct subcellular autophagy impairments in induced neurons from patients with Huntington's disease. Brain, 145(9), 3035-3057.

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Mitochondrial Dysfunction and Autophagy Modulation in Chondrocytes

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TC28a2 cells and normal human chondrocytes were plated in 8-well chamber slides (1×10⁵ cells), 6-well plates (5×10⁵ cells) or 12-well (2.5×10⁵ cells) plates, in DMEM containing 2% fetal calf serum (FCS, Life Technologies, NY, USA). Small molecule mitochondrial respiratory chain inhibitor (Oligomycin; 10 μg/ml, Sigma-Aldrich, St. Louis, MO, USA) or autophagy activators (Rapamycin; 10 μM, Calbiochem and Torin 1 50 nM, Tocris Biosciences, UK) were added and incubated for the indicated times. Antimycin A; 40 μg/ml, Valinomycin; 1 μM, H₂O₂; 100 μM and TNFα (10 ng/ml) + Actinomycin D (1 μg/ml) (Sigma-Aldrich) were employed as positive controls for mitochondrial dysfunction, mitochondrial membrane depolarization, ROS production and cell death, respectively.
For siRNA transfection experiments, normal human chondrocytes were plated in a 6-well plates (1.5×10⁵ cells) in DMEM containing 10% FCS without antibiotics overnight before transfection.

de Figueroa P.L., Lotz M., Blanco F.J, & Caramés B. (2015). Autophagy Activation Protects from Mitochondrial Dysfunction in Human Chondrocytes. Arthritis & rheumatology (Hoboken, N.J.), 67(4), 966-976.

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Cellular Response to Doxorubicin and mTOR Inhibitors

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HeLa and ARPE-19 cells were purchased from ATCC. ARPE-19 is a spontaneously immortalized cell line of human retinal pigment epithelium. These cells are diploid and non-transformed. HeLa TFEB and TFEB/TFE3 knockout and p53-null human osteosarcoma (SAOS-2 p53-null) cells were kindly provided by Richard J. Youle (National Institutes of Health, Bethesda, MD, USA) and Dr. Caterina Missero (Centro di Ingegneria Genetica e Biotecnologie Avanzate, Naples, Italy), respectively. HeLa cells stably expressing TFEB-GFP or TFEB 3xflag were obtained as previously described [22 (link),28 (link)]. Cells were cultured at 37 °C and 5% CO2 in DMEM (Sigma-Aldrich, St. Louis, MO, USA, D6429) supplemented with fetal bovine serum (10%) and Pen/Strep (1%). Treatments with doxorubicin (D1515, Sigma, St. Louis, MO, USA, D6429), Torin (Tocris, Bristol, UK), or UC2288 (ab146969, Abcam, Cambridge, UK) were performed at indicated time points and concentrations and compared with control DMSO. For colony formation assay, about 200 cells per well were seeded in a 6-well plate. Next day cells were treated, and maintained in culture 8 days more, replacing medium every three days.

Pisonero-Vaquero S., Soldati C., Cesana M., Ballabio A, & Medina D.L. (2020). TFEB Modulates p21/WAF1/CIP1 during the DNA Damage Response. Cells, 9(5), 1186.

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Partial Liver Warm Ischemia-Reperfusion Injury Model

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A model of partial liver warm IRI was used (51 (link)). In brief, an atraumatic clip was used to interrupt the arterial and portal venous blood supply to the cephalad lobes of the liver for 90 min. Sham controls underwent the same procedure, but without vascular occlusion. Rapamycin (1-5mg/kg) or Torin 1(10mg/kg), or chloroquine (CQ, 60mg/kg) (Tocris Bioscience, Cambridge, UK) or 3-Methyladenine (3-MA, 30mg/kg), or Triciribine (1mg/kg) (Sigma, St. Luis, MO), were administered, i.p., 1 h prior to the onset of liver ischemia.
Mice were sacrificed after 6h of reperfusion. Serum alanine aminotransferase (sALT) levels were measured by IDEXX Laboratories. Part of liver specimens were fixed in 10% buffered formalin and embedded in paraffin. Liver sections (4μm) were stained with hematoxylin and eosin, and then analyzed blindly. The severity of liver IRI was graded blindly using Suzuki's criteria on a scale from 0 to 4 (52 (link)).

Zhu J., Lu T., Yue S., Shen X., Gao F., Busuttil R.W., Kupiec-Weglinski J.W., Xia Q, & Zhai Y. (2015). Rapamycin protects livers from ischemia and reperfusion Injury via both autophagy induction and mTORC2-Akt activation. Transplantation, 99(1), 48-55.

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Cytokine Profiling of Stimulated PBMCs

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For stimulation experiments, 5 × 10⁶ PBMCs/mL or 1 × 10⁶ monocytes/mL were stimulated with RPMI, 1 μg/mL Mtb strain H37Rv (Mtb) lysate, 10 μg/mL Pam3Cys (EMC Microcollections, Germany) and/or 10 μg/mL MDP (Sigma) in the presence or absence of 1 or 10 nM rapamycin (LC‐laboratories), 100 nM Torin (Tocris), 50 or 500 μM sodium l‐ascorbate (Sigma), 50 or 500 μM AICAR (Brunschwig chemie), 100 nM wortmannin (Cayla Invivogen), or 20 or 100 ng/mL of TLR4 inhibitory ligand double‐extracted B. quintana LPS for 24 h or with 10% pooled human serum for 7 days. PBMCs from patients with the homozygous carriage of a frameshift in Nod2 due to an insertion of cysteine at position 1007 (1007finsC) (rs2066847) were stimulated with RPMI, MDP, or Mtb for 1 or 7 days. Culture supernatants were collected and stored at −20°C. For Western blots, 5 × 10⁶ PBMCs were stimulated with RPMI, 1 μg/mL Mtb lysate or 50 ng/mL GM‐CSF in the presence or absence of the aforementioned inhibitors. Cytokine measurements from cell culture supernatants were performed by ELISA namely IL‐1β, TNF‐α, IL‐17A, IL‐22 (R&D Systems, Minneapolis, MN); and IL‐6, IFN‐γ, and IL‐10 (Sanquin, Amsterdam) were measured.

Lachmandas E., Beigier‐Bompadre M., Cheng S., Kumar V., van Laarhoven A., Wang X., Ammerdorffer A., Boutens L., de Jong D., Kanneganti T., Gresnigt M.S., Ottenhoff T.H., Joosten L.A., Stienstra R., Wijmenga C., Kaufmann S.H., van Crevel R, & Netea M.G. (2016). Rewiring cellular metabolism via the AKT/mTOR pathway contributes to host defence against Mycobacterium tuberculosis in human and murine cells. European Journal of Immunology, 46(11), 2574-2586.

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Signaling Pathways in Keratinocyte Differentiation

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All chemicals were purchased from Sigma unless stated otherwise. Rapamycin, Wortmannin, LY294002 and MHY1485 were from Calbiochem. Torin was purchased from Tocris Bioscience. Cytokines were obtained from Peprotech.
Phospho-specific (P-mTOR S2448 #5536, P-mTOR S2448 #2976, P-PRAS T246 #2997, P-S6 S235/6 #2211, P-Akt S473 #4060, P-ERk1 T202/Y204 #4370, P-p38MAPK T180/Y182 #4511, and corresponding pan antibodies (PRAS #2691, Akt #2938, S6 #2217) Rheb antibody (AP53656PU-N) was from Acris and tubulin #2128 antibody was from Cell Signaling Technology. Raptor antibody (sc27744) was from Santa Cruz, actin antibody (A1978) was from Sigma, involucrin antibody (ab20202) and β1-integrin antibody (ab30394) were from Abcam and filaggrin (PRB-417P-100) was from Convance. Ki-67 (MB67) antibody was obtained from Novus Biologicals and Keratin6 antibody (Ks6.KA12) from Thermo Scientific. Involucrin (Poly19244) for mouse IHC was from Biolegend.

Buerger C., Shirsath N., Lang V., Berard A., Diehl S., Kaufmann R., Boehncke W.H, & Wolf P. (2017). Inflammation dependent mTORC1 signaling interferes with the switch from keratinocyte proliferation to differentiation. PLoS ONE, 12(7), e0180853.

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Signaling Pathway Modulation Protocol

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SB2035800 (p38 inhibitor, Selleck Chemical), PD0325901 (MEK1/2 inhibitor, Sigma), Torin (mTOR inhibitor, Tocris Bioscience), cycloheximide (translation inhibitor, Sigma), and actinomycin D (transcription inhibitor, Sigma) were re-suspended in DMSO.

Cicchetto A.C., Jacobson E.C., Sunshine H., Wilde B.R., Krall A.S., Jarrett K.E., Sedgeman L., Turner M., Plath K., Luisa Iruela-Arispe M., de Aguiar Vallim T.Q, & Christofk H.R. (2023). ZFP36-mediated mRNA decay regulates metabolism. Cell reports, 42(5), 112411.

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Signaling Pathway Modulation Protocol

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Immunoblotting of Kinase Pathway Markers

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Antibodies to p-S6K Thr389 (9205), S6K (9202), p-AKT Ser473 (9271), AKT (2920), p-S6 Ser240/4 (2215), S6 (2317), p-PKD Ser744/8 (2054) and p-PKC substrate (2261) were from Cell Signaling (Danvers, MA, USA), anti-PKCα Ser657 from US Biologicals (Swappscott, MA, USA) (P9103-16A), anti-PKCα and PKD from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (sc-208, -935, -935), anti-SREBP-1 from Millipore (Temecula, CA, USA) (04-469), anti-DGKζ from Abcam (Cambridge, UK) (ab105195), anti-DGKα from Abnova (Tapei, Taiwan) (BO1P), anti-tubulin DM1A (T9026) from Sigma-Aldrich (Saint Louis, MO, USA) and anti-phosphotyrosine (4G10) from Upstate (Lake Placid, NY, USA). Horseradish peroxidase-coupled polyclonal goat anti-mouse and -rabbit immunoglobulins were from DakoCytomation (Glostrup, Denmark) (P0447, P0448). DGK inhibitor II (R59949; 266788), bisindolylmaleimide and Gö6976 were from Calbiochem (Darmstadt, Germany), (203291, 36520). Rapamycin was from LC Laboratory (Woburn, MA, USA) (R-5000). Torin was from Tocris (Bristol, UK) (4247). Recombinant DGK from Escherichia coli (D3065), PMSF, leupeptin and aprotinin were from Sigma-Aldrich. Adenosine 5′triphosphate [γ³²P] was from Hartman Analytic (Braunschweig, Germany). All lipids were purchased from Avanti Polar Lipids (Alavaster, AL, USA). Organic reagents were from Merck (Darmstadt, Germany).

Torres-Ayuso P., Tello-Lafoz M., Mérida I, & Ávila-Flores A. (2015). Diacylglycerol kinase-ζ regulates mTORC1 and lipogenic metabolism in cancer cells through SREBP-1. Oncogenesis, 4(8), e164-.

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