Akti 1 2

Manufactured by Bio-Techne

Sourced in United Kingdom

Akti-1/2 is a laboratory instrument designed for the detection and quantification of kinase activity. It utilizes a fluorescence-based assay to measure the phosphorylation of a specific substrate by a target kinase. The core function of Akti-1/2 is to provide researchers with a reliable tool for studying kinase activity in a controlled laboratory setting.

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Lab products found in correlation

4 protocols using akti 1 2

Small Molecule Inhibitor Protocol

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Sodium salicylate (CAS N 54-21-7), oligomycin from Streptomyces diastatochromogenes (CAS N 1404-19-9) and doxycycline monohydrate (CAS N 17086-28-1) were purchased from Sigma-Aldrich (St. Louis, MO, USA). 17-DMAG (CAS N150270-08-9) was purchased from Tebu-bio (Barcelona, Spain). AICAR (CAS N 2627-69-2), Akti-1/2 (CAS N 612847-09-3), dorsomorphin dihydrochloride (CAS N 1219168-18-9), SC79 (CAS N 305834-79-1), and (R)-(+)-etomoxir sodium salt (CAS N 828934-41-4) were purchased from TOCRIS (Bristol, UK).
AICAR, Akti-1/2, dorsomorphin, etomoxir, and SC79 stock dilutions were prepared in DMSO at 20 mM whereas oligomycin and 17-DMAG were prepared at 2 and 1 mmol L⁻¹, respectively. The drug Sodium salicylate was prepared before use in the appropriate culture media at a starting concentration of 200 µmol L⁻¹.

Martinez-Peinado N., Martori C., Cortes-Serra N., Sherman J., Rodriguez A., Gascon J., Alberola J., Pinazo M.J., Rodriguez-Cortes A, & Alonso-Padilla J. (2021). Anti-Trypanosoma cruzi Activity of Metabolism Modifier Compounds. International Journal of Molecular Sciences, 22(2), 688.

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Hepatic and Cellular Triglyceride Measurement

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Hepatic triglyceride content was assessed according to Carr et al. [24 (link)]. For the evaluation of cellular triglyceride content, cells were grown in monolayer, resuspended in PBS with 0.5% Triton X-100 and lysed with an ultrasound tip for evaluation with a commercial kit (Zen-bio, USA), according to Beller M. (personal communication). Compounds used were murine or human recombinant Igfbp3 (R&D systems, 0.3 μg/mL), Cytochalasin D (Sigma-Aldrich, 1:1000), PP2 (Sigma-Aldrich, 10 uM), recombinant SerpinA12 (R&Dsystems, 100 ng/mL), Akt inhibitor (Tocris, Akti-1/2, 250 nM), PQ-401 (Tocris, 10uM) and AMPK inhibitor (Tocris, Dorsomorphin dihydrochloride, 20 uM).

Arab J.P., Cabrera D., Sehrawat T.S., Jalan-Sakrikar N., Verma V.K., Simonetto D., Cao S., Yaqoob U., Leon J., Freire M., Vargas J.I., De Assuncao T.M., Kwon J.H., Guo Y., Kostallari E., Cai Q., Kisseleva T., Oh Y., Arrese M., Huebert R.C, & Shah V.H. (2020). Hepatic stellate cell activation promotes alcohol-induced steatohepatitis through Igfbp3 and SerpinA12. Journal of hepatology, 73(1), 149-160.

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Modulating mGlu5 Signaling Pathways

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LY379268 (200 nM), (S)-DHPG (100 μM), and TTX (500 nM) were purchased from Abcam (Cambridge, U.K.). EGTA (20 mM) and BAPTA (20 mM) were purchased from Sigma. Carbachol (100 μM), apamin (100 nM), LY294002 (20 μM), Akti-1/2 (10 μM), KU-0063794 (1 μM), anisomycin (20 μM), and CHIR 99021 (2 μM) were purchased from Tocris (Bristol, U.K.). VU0409551 (10 μM) and VU0469650 (10 μM) were synthesized in-house. The control, mutated peptide mGlu₅-mut (YGRKKRRQRRRALTPLSPRR) and the active, dominant negative mGlu₅-C-ter (YGRKKRRQRRRALTPPSPFR) (Mao et al., 2005 (link); Ronesi and Huber, 2008 (link)) were prepared by Bio-Synthesis (Lewisville, TX, U.S.A.) and added to the internal solution at 20 nM.

Joffe M.E., Santiago C.I., Stansley B.J., Maksymetz J., Gogliotti R.G., Engers J.L., Nicoletti F., Lindsley C.W, & Conn P.J. (2018). Mechanisms underlying prelimbic prefrontal cortex mGlu3/mGlu5-dependent plasticity and reversal learning deficits following acute stress. Neuropharmacology, 144, 19-28.

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Modeling Cardiomyopathy Pathogenesis with Engineered hiPSC-CMs

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The control human iPSC line (Stanford Cardiovascular Institute [SCVI-113]) was obtained from the Stanford CVI iPSC Biobank. The P710R mutation was edited into these cells using a method that has been described previously (83 (link), 84 (link)). Cells were differentiated and their contractile function was assessed on micropatterned hydrogels gels as previously described (35 (link), 37 (link)). Cells on micropatterened gels were fixed and stained for β-cardiac myosin and their sarcomere length quantifed. Cells were also plated onto micropatterned film and imaged with TEM. Unpatterned cells were stained for cardiac troponin T to measure cell area. To clarify the effect of ERK and Akt, the cells were treated with either Akt inhibitor (Akti 1/2; Tocris; 5 µM) or and ERK inhibitor (SCH772984; SelleckChem; 1 µM) every 3 d between day 26 and day 46 (a time frame over which hiPSC-CMs have been shown to increase in area). Additional details are included in the SI Appendix.

Vander Roest A.S., Liu C., Morck M.M., Kooiker K.B., Jung G., Song D., Dawood A., Jhingran A., Pardon G., Ranjbarvaziri S., Fajardo G., Zhao M., Campbell K.S., Pruitt B.L., Spudich J.A., Ruppel K.M, & Bernstein D. (2021). Hypertrophic cardiomyopathy β-cardiac myosin mutation (P710R) leads to hypercontractility by disrupting super relaxed state. Proceedings of the National Academy of Sciences of the United States of America, 118(24), e2025030118.

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