Akt inhibitor x

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Akt inhibitor X is a chemical compound that inhibits the activity of the Akt (Protein Kinase B) signaling pathway. Akt is a key regulator of cellular processes such as cell growth, proliferation, and survival. Akt inhibitor X can be used in research applications to investigate the role of the Akt pathway in various biological systems.

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Lab products found in correlation

10 protocols using akt inhibitor x

Pharmacological Inhibition of mTOR Pathway

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Rapamycin (LC Laboratories, Woburn, MA, USA), PP242 and Akt inhibitor X (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were dissolved in dimethyl sulfoxide (DMSO) to prepare 100 μg/ml Rapamycin, 1 mM PP242 and 10 mM Akt inhibitor X as stock solutions, and stored at −20°C. IGF-1 (PeproTech, Rocky Hill, NJ, USA) was rehydrated in 0.1 M acetic acid to prepare a 10 μg/ml stock solution and stored at −80°C. Bovine collagen type IV (CN IV) was purchased from Rockland Immunochemicals (Gilbertsville, PA, USA). 7-methyl GTP Sepharose 4B was from GE Healthcare Life Sciences (Piscataway, NJ, USA). CellTiter 96^® AQ_ueous One Solution Cell Proliferation Assay Kit was from Promega (Madison, WI, USA). Enhanced chemiluminescence solution was from Pierce (Rockford, IL, USA). Antibodies included those against S6K1, Akt, S6, eIF4E, GSK3β, HA (Santa Cruz Biotechnology, Santa Cruz, CA, USA), mTOR, phospho-S6K1 (Thr389), phospho-Akt (Ser473), phospho-Akt (Thr308), phospho-S6 (Ser/235/236), 4E-BP1, phospho-4E-BP1 (Thr70) (Cell Signaling, Beverly, MA, USA), raptor, rictor (Bethyl Laboratories, Montgomery, TX, USA), phospho-GSK3β (Ser9) (Epitomics, Burlingame, CA, USA), β-tubulin, FLAG (Sigma, St. Louis, MO, USA), goat anti-rabbit IgG-horseradish peroxidase (HRP), goat anti-mouse IgG-HRP, and rabbit anti-goat IgG-HRP (Pierce). All other chemicals were purchased from Sigma.

Chen L., Xu B., Liu L., Liu C., Luo Y., Chen X., Barzegar M., Chung J, & Huang S. (2015). Both mTORC1 and mTORC2 are involved in the regulation of cell adhesion. Oncotarget, 6(9), 7136-7150.

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Antioxidant and Cell Signaling Pathway Modulation

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Resveratrol, cadmium chloride, poly-d-lysine (PDL), U0126, SP600125 and protease inhibitor cocktail were purchased from Sigma (St Louis, MO, USA). Rapamycin was bought from ALEXIS (San Diego, CA, USA), whereas 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate (CM-H2DCFDA) was from MP Biomedicals (Solon, OH, USA). PP242 and Akt inhibitor X were provided by Santa Cruz Biotechnology (Santa Cruz Biotechnology, Dallas, TX, USA). Dulbecco’s modified Eagle medium (DMEM), 0.05% Trypsin-EDTA, NEURO-BASAL™ Media, and B27 Supplement were purchased from Invitrogen (Grand Island, NY, USA). Horse serum and fetal bovine serum (FBS) were supplied by Hyclone (Logan, UT, USA). Enhanced chemiluminescence solution was from Sciben Biotech Company (Nanjing, China). Other chemicals used in this work were obtained from local commercial sources and were of analytical grade.

Liu C., Zhang R., Yang L., Ji T., Zhu C., Liu B., Zhang H., Xu C., Zhang N., Huang S, & Chen L. (2022). Neuroprotection of resveratrol against cadmium-poisoning acts through dual inhibition of mTORC1/2 signaling. Neuropharmacology, 219, 109236.

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Maduramicin and Akt Inhibitor X Protocol

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Maduramicin ammonium and Akt inhibitor X were provided by Santa Cruz Biotechnology (Santa Cruz, CA, USA), whereas chloroquine diphosphate (CQ), monodansylcadaverine (MDC), 4′,6-diamidino-2-phenylindole (DAPI) and protease inhibitor cocktail were purchased from Sigma (St Louis, MO, USA). Rapamycin were from ALEXIS Biochemicals Corporation (San Diego, CA, USA). Dulbecco’s modified Eagle medium (DMEM) and 0.05% Trypsin-EDTA were purchased from Invitrogen (Grand Island, NY, USA). Fetal bovine serum (FBS) was supplied by Hyclone (Logan, UT, USA). Enhanced chemiluminescence solution was from Sciben Biotech Company (Nanjing, China). Other chemicals were purchased from local commercial sources and were of analytical grade.

Dong X., Zhao R., Li Y., Yu Q., Chen X., Hu X., Ma J., Chen X., Huang S, & Chen L. (2019). Maduramicin inactivation of Akt impairs autophagic flux leading to accumulated autophagosomes-dependent apoptosis in skeletal myoblast cells. The international journal of biochemistry & cell biology, 114, 105573.

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Flow Cytometry Assay for Lymphocyte Activation

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Anti-CD19 magnetic fluorobeads-B was purchased from One Lambda (Canoga Park, CA, USA). PP242 (mTORC1/2 kinase inhibitor) and Akt inhibitor X were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rapamycin (mTOR inhibitor) was from ALEXIS (San Diego, CA, USA). Refolded human soluble BAFF (hsBAFF) was a recombinant form of the extracellular domain of the BAFF produced in Escherichia coli from this group (Cao et al., 2005 (link)). RPMI 1640 was from Gibco (Rockville, MD, USA). Fetal bovine serum (FBS) was supplied by Hyclone (Logan, UT, USA). CellTiter 96^® AQ_ueous One Solution Cell Proliferation Assay kit was from Promega (Madison, WI, USA). Other chemicals were purchased from local commercial sources and were of analytical grade.

Zeng Q., Qin S., Zhang H., Liu B., Qin J., Wang X., Zhang R., Liu C., Dong X., Zhang S., Huang S, & Chen L. (2017). Rapamycin attenuates BAFF-extended proliferation and survival via disruption of mTORC1/2 signaling in normal and neoplastic B-lymphoid cells. Journal of cellular physiology, 233(1), 516-529.

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Anti-CD19 Magnetic Fluorobeads-B Assay

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Anti-CD19 magnetic fluorobeads-B was purchased from One Lambda (Canoga Park, CA, USA). Akt inhibitor X and PP242 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Refolded human soluble BAFF (hsBAFF) was a recombinant form of the extracellular domain of the BAFF produced in Escherichia coli from this group [35 (link)]. RPMI 1640 medium and fetal bovine serum (FBS) were supplied by Gibco (Rockville, MD, USA). CellTiter 96®AQ_ueous One Solution Cell Proliferation Assay kit was from Promega (Madison, WI, USA). Enhanced chemiluminescence solution was from Sciben Biotech Company (Nanjing, China). 1,2-bis(oAminophenoxy) ethane-N,N,N’,N’-tetraacetic acid tetra (acetoxymethyl) ester (BAPTA/AM) and 2-aminoethoxydiphenyl borane (2-APB) were purchased from Calbiochem (San Diego, CA, USA), whereas ethylene glycol tetra-acetic acid (EGTA) was purchased from Sigma (St. Louis, MO, USA). ML-SA1 was supplied by Abcam Technology (Cambridge, UK). Verapamil hydrochloride was from MedChemExpress (Monmouth Junction, NJ, USA). Rapamycin and KN93 were from ALEXIS (San Diego, CA, USA), while U0126 was from Sigma. Other chemicals were purchased from local commercial sources and were of analytical grade.

Zeng Q., Zhou Z., Qin S., Yao Y., Qin J., Zhang H., Zhang R., Xu C., Zhang S., Huang S, & Chen L. (2020). Rapamycin inhibits B-cell activating factor (BAFF)-stimulated cell proliferation and survival by suppressing Ca2+-CaMKII-dependent PTEN/Akt-Erk1/2 signaling pathway in normal and neoplastic B-lymphoid cells. Cell calcium, 87, 102171.

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Investigating mTOR Signaling Pathway

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Cadmium chloride, poly-D-lysine (PDL), 4′,6-diamidino-2-phenylindole (DAPI), and protease inhibitor cocktail were purchased from Sigma (St Louis, MO, USA). Rapamycin was from ALEXIS (San Diego, CA, USA). Dulbecco’s modified Eagle medium (DMEM), 0.05% Trypsin-EDTA, NEUROBASAL^™ Media, and B27 Supplement were purchased from Invitrogen (Grand Island, NY, USA). Horse serum and fetal bovine serum (FBS) were supplied by Hyclone (Logan, UT, USA). Enhanced chemiluminescence solution was from Millipore (Billerica, MA, USA). Akt inhibitor X was provided by Santa Cruz Biotechnology (Santa Cruz, CA, USA). The following antibodies were used: phospho-Akt (Ser473), 4E-BP1, phospho-4E-BP1 (Thr70), S6 ribosomal protein, phospho-S6 ribosomal protein (Ser235/236), phospho-S6K1 (Thr389), caspase-3, and PARP (all from Cell Signaling Technology, Beverly, MA, USA); Akt, GSK3β, and S6K1 (all from Santa Cruz Biotechnology); phospho-GSK3β (Ser9) (Epitomics, Burlingame, CA, USA); raptor and rictor (Bethyl Laboratories, Montgomery, TX, USA); HA, mTOR, phospho-Akt (Thr308), and β-tubulin (all from Sigma); goat anti-rabbit IgG-horseradish peroxidase (HRP), goat anti-mouse IgG-HRP, and rabbit anti-goat IgG-HRP (Pierce, Rockford, IL, USA). Other chemicals were purchased from local commercial sources and were of analytical grade.

Xu C., Liu C., Liu L., Zhang R., Zhang H., Chen S., Luo Y., Chen L, & Huang S. (2015). Rapamycin prevents cadmium-induced neuronal cell death via targeting both mTORC1 and mTORC2 pathways. Neuropharmacology, 97, 35-45.

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Evaluating Autophagy Modulation in Cells

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Cadmium chloride, poly-D-lysine (PDL), 3-methyladenine (3-MA), chloroquine diphosphate (CQ), monodansylcadaverine (MDC), 4′,6-diamidino-2-phenylindole (DAPI), and protease inhibitor cocktail were purchased from Sigma (St Louis, MO, USA), whereas Akt inhibitor X was provided by Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rapamycin were from ALEXIS Biochemicals Corporation (San Diego, CA, USA). Dulbecco’s modified Eagle medium (DMEM), 0.05% Trypsin-EDTA, NEUROBASALTM Media, and B27 Supplement were purchased from Invitrogen (Grand Island, NY, USA). Horse serum and fetal bovine serum (FBS) were supplied by Hyclone (Logan, UT, USA). Other chemicals were purchased from local commercial sources and were of analytical grade.

Zhang H., Dong X., Zhao R., Zhang R., Xu C., Wang X., Liu C., Hu X., Huang S, & Chen L. (2018). Cadmium results in accumulation of autophagosomes-dependent apoptosis through activating Akt-impaired autophagic flux in neuronal cells. Cellular signalling, 55, 26-39.

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Resveratrol-Modulated Extracellular Matrix Signaling

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Resveratrol, collagen type IV (CN IV), fibronectin, laminin, U0126 and okadaic acid were purchased from Sigma (Saint Louis, MO, USA). Resveratrol was dissolved in dimethyl sulfoxide (DMSO) to prepare a stock solution (100 mM), aliquoted and stored at −80˚C. IGF-1 was from PeproTech (Rocky Hill, NJ, USA), rehydrated in 0.1 M acetic acid to prepare a 10 μg/ml stock solution and stored at −80°C. RPMI 1640 medium, Dulbecco’s modified Eagle’s medium (DMEM) and 0.05% trypsin-EDTA were obtained from Invitrogen (Grand Island, NY, USA). Fetal bovine serum (FBS) was from Hyclone (Logan, UT, USA). Akt inhibitor X was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rapamycin was purchased from LC Laboratories (Woburn, MA, USA). CellTiter 96^® AQ_ueous One Solution Cell Proliferation Assay Kit was from Promega (Madison, WI, USA). Enhanced chemiluminescence solution was from Millipore (Billerica, MA, USA). Other chemicals were purchased from local commercial sources and were of analytical grade.

Chen X., Hu X., Li Y., Zhu C., Dong X., Zhang R., Ma J., Huang S, & Chen L. (2018). Resveratrol inhibits Erk1/2-mediated adhesion of cancer cells via activating PP2A/PTEN signaling network. Journal of cellular physiology, 234(3), 2822-2836.

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Signaling Pathways Modulation in B Cells

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Anti-CD19 magnetic fluorobeads-B was purchased from One Lambda (Canoga Park, CA, USA). Metformin, U0126 and protease inhibitor cocktail were purchased from Sigma (St Louis, MO, USA), whereas Akt inhibitor X were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Rapamycin was from ALEXIS (San Diego, CA, USA). Refolded human soluble BAFF (hsBAFF) was a recombinant form of the extracellular domain of the BAFF produced in Escherichia coli from this group [34 (link)]. RPMI 1640 medium and fetal bovine serum (FBS) were from Gibco (Rockville, MD, USA). CellTiter 96®AQ_ueous One Solution Cell Proliferation Assay kit was from Promega (Madison, WI, USA). Enhanced chemiluminescence solution was supplied by Sciben Biotech Company (Nanjing, China). The following primary antibodies were used: phosphorylated Akt (p-Akt) (Ser473), p-S6K1 (Thr389), p-S6 (Ser235/236) (Cell Signaling Technology, Danvers, MA, USA), S6K1, S6, p-Erk1/2 (Thr202/Tyr204), Erk2 (Santa Cruz Biotechnology), p-PTEN (Thr366), PTEN (Epitomics, Burlingame, CA, USA), MKK1, mTOR (Sigma), Erk1/2, survivin, Akt, β-actin, FLAG, HA (Sciben Biotech Company); Goat anti-rabbit IgG-horseradish peroxidase (HRP), goat anti-mouse IgG-HRP, and rabbit anti-goat IgG-HRP (Sciben Biotech Company). Other chemicals used in this work were purchased from local commercial sources and were of analytical grade.

Chen X., Ma J., Yao Y., Zhu J., Zhou Z., Zhao R., Dong X., Gao W., Zhang S., Huang S, & Chen L. (2021). Metformin prevents BAFF activation of Erk1/2 from B-cell proliferation and survival by impeding mTOR-PTEN/Akt signaling pathway. International immunopharmacology, 96, 107771.

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Molecular Signaling Pathway Analysis

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Cadmium chloride, 4′, 6-diamidino-2-phenylindole (DAPI), poly-D-lysine (PDL), Okadaic acid, PD98059, and protease inhibitor cocktail were purchased from Sigma (St Louis, MO, USA). Rapamycin was from ALEXIS (San Diego, CA, USA). Dulbecco's modified Eagle medium (DMEM), 0.05% Trypsin-EDTA, NEUROBASAL™ Media, and B27 Supplement were purchased from Invitrogen (Grand Island, NY, USA). Horse serum and fetal bovine serum (FBS) were supplied by Hyclone (Logan, UT, USA). Enhanced chemiluminescence solution was from Millipore (Billerica, MA, USA). Akt inhibitor X was provided by Santa Cruz Biotechnology (Santa Cruz, CA, USA). The following antibodies were used: PP2ACα (BD Biosciences, San Jose, CA, USA); phospho-S6K1 (Thr389), phospho-Akt (Ser473), and cleaved-caspase-3 (Cell Signaling Technology, Beverly, MA, USA); demethylated-PP2A, Akt, phospho-Erk1/2 (Thr202/Tyr204), Erk2, and S6K1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); phospho-PP2A, and PTEN (Epitomics, Burlingame, CA, USA); phospho-Akt (Thr308), FLAG, HA, mTOR, and β-tubulin (all from Sigma); goat anti-rabbit IgG-horseradish peroxidase (HRP), goat anti-mouse IgG-HRP, and rabbit anti-goat IgG-HRP (Pierce, Rockford, IL, USA). Other chemicals were purchased from local commercial sources and were of analytical grade.

Xu C., Zhang H., Liu C., Zhu Y., Wang X., Gao W., Huang S, & Chen L. (2015). Rapamycin inhibits Erk1/2-mediated neuronal apoptosis caused by cadmium. Oncotarget, 6(25), 21452-21467.

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