Phospho akt

Manufactured by Cell Signaling Technology

Sourced in United States, United Kingdom, China, Canada, Germany, France

Phospho-Akt is a lab equipment product that detects and quantifies phosphorylated Akt protein. It is used to study the activation status of the Akt signaling pathway.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Anti-phospho-AKT-substrate (23C8D2), Phospho-AKT (Thr308) Phospho-Akt Pathway Antibody Sampler Kit Phospho-Akt rabbit mAb Phospho-AKT (4060P, Anti-Phospho-Akt (Thr308) clone C31E5E Phospho-AKT (Serine 473), Anti-phospho-Akt XP Anti-AKT and anti-Phospho-AKT-Ser473 Anti-phospho-Akt (9271)

892 protocols using phospho akt

Comprehensive Immunoblotting Analysis of mTOR Pathway

Check if the same lab product or an alternative is used in the 5 most similar protocols

All the antibodies were from Cell Signaling Technology (Danvers, MA, USA): mTOR (cat #2983), phospho-mTOR (Ser2481, cat #2974), phospho-mTOR (Ser2448, cat #5536), Rictor (cat #9476), phospho-Rictor (Thr1135, cat #3806), Raptor (cat #2280), AKT (cat #4691), phospho-AKT (Ser473, cat #4060), phospho-AKT (Thr308, cat #13038), phospho-AKT (Thr450, cat #9267), GSK-3β (cat #9315), phospho-GSK-3β (Ser9, cat #9336), PTEN (cat #9559), β-actin (cat #4970), 4E-BP1 (cat #9644), phospho-4E-BP1 (Thr37/46, cat #2855), S6K1/p70S6K (cat #9202), phospho-S6K1/phospho-p70S6K (Thr389, cat #9234), Myc-Tag (cat #2276), horseradish peroxidase-linked anti-rabbit IgG (cat #7074), horseradish peroxidase-linked anti-mouse IgG (cat #7076). Lipofectamine LTX, plus reagent, culture medium were from Invitrogen (Carlsbad, CA, USA). Antibiotic–antimycotic, propedium iodide (PI), rapamycin, GSK3β inhibitor (SB212763), PI3K inhibitor (LY294002 and wortmannin) and other chemicals were from Sigma–Aldrich (St Louis, MO, USA). Protease and phosphatase inhibitor cocktails were from Calbiochem (San Diego, CA, USA). Cycle Test Plus kit was from BD Bioscience (East Rutherford, NJ, USA). Super Signal West Pico imaging system was from Thermo Scientific (Rockford, IL, USA).

Bhattacharya K., Maiti S, & Mandal C. (2016). PTEN negatively regulates mTORC2 formation and signaling in grade IV glioma via Rictor hyperphosphorylation at Thr1135 and direct the mode of action of an mTORC1/2 inhibitor. Oncogenesis, 5(5), e227-.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

To prepare whole-cell lysates, cells were washed twice with cold PBS and lysed in M-PER Mammalian Protein Extraction Reagent (product no. 78501, Thermo Fisher Scientific) supplemented with Pierce protease and phosphatase inhibitor cocktail (product no. A32961, Thermo Fisher Scientific). Tumor tissue samples were lysed in T-PER Tissue Protein Extraction Reagent (product no. 78510, Thermo Fisher Scientific) with above supplement. The automatic hand-operated OMNI-TIP Homogenizer (Omni International, Inc.) was used to homogenize the tumor tissues. Lysates from whole cells and tumor homogenates were then processed for SDS-PAGE Western blotting. The following antibodies were used: Phospho-MEK1/2 (S217/221; catalog no. 9154S), MEK1/2 (catalog no. 4694S), Phospho-AKT (S473; catalog no. 9271S), Phospho-AKT (T308; catalog no. 4056S), AKT (catalog no. 9272S), Phospho-Erk1/2 (catalog no. 4370S), Erk1/2 (catalog no. 4695S), Phospho-S6 Ribosomal Protein (Ser 235/236; catalog no. 4858S), S6 Ribosomal Protein (catalog no.2217S), cleaved-CASP-3 (catalog no. 9664), cleaved-PARP (catalog no. 9541) from Cell Signaling Technology; anti-β-ACTIN (catalog no A5441-.2ML from Sigma-Aldrich; KRAS (catalog no. OP24, from EMDMillipore), and RAF1 (catalog no. sc-7267, Santa Cruz Biotechnology), horseradish peroxidase–conjugated goat anti-GST antibody (catalog no. A190-121P from Bethyl Laboratories).

Kazi A., Ranjan A., Kumar M.V. V., Agianian B., Garcia Chavez M., Vudatha V., Wang R., Vangipurapu R., Chen L., Kennedy P., Subramanian K., Quirke J.C., Beato F., Underwood P.W., Fleming J.B., Trevino J., Hergenrother P.J., Gavathiotis E, & Sebti S.M. (2023). Discovery of KRB-456, a KRAS G12D Switch-I/II Allosteric Pocket Binder That Inhibits the Growth of Pancreatic Cancer Patient-derived Tumors. Cancer Research Communications, 3(12), 2623-2639.

+ Open protocol

+ Expand

Comprehensive Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

AR(rabbit monoclonal, 1:1000, #5153), PSA/KLK3(rabbit monoclonal, 1:1000, #5365), TOMM20(rabbit monoclonal, 1:1000, #42,406), ATG5(rabbit monoclonal, 1:1000, #12994S), LC3I/II(rabbit monoclonal, 1:1000, #12,741), Akt (rabbit monoclonal, 1:1000, #4691), Phospho-Akt (Ser473)(rabbit monoclonal, 1:1000, #4060), Phospho-Akt (Thr308)(rabbit monoclonal, 1:1000, #13,038), Nanog(rabbit monoclonal, 1:1000, #4903), Sox2(rabbit monoclonal, 1:1000, #3579), ALDH1A1(rabbit monoclonal, 1:1000, #36671S) and VDAC(rabbit monoclonal, 1:1000, #4661) antibodies were purchased from Cell Signaling Technology (CST). NCAM1(rabbit polyclonal antibody, 1:1000, A7913), β-Actin(mouse polyclonal, 1:5000, AC004) and α-Tublin(mouse polyclonal, 1:5000, AC012) antibodies were purchased from ABclonal Technology (Upper Heyford, UK). NSE (rabbit monoclonal, 1:1000, ab180943) and Syp (Rabbit monoclonal, 1:1000, ab184176) were purchased from Abcam (Shanghai, China). ALDH1A1(mouse monoclonal antibody, 1:500,sc-166362), TOMM70(mouse monoclonal antibody, 1:500, sc-390545),AR(rabbit polyclonal, 1:1000, Sc-815X) antibodies were purchased from Santa cruz. E-cadherin (rabbit polyclonal, 1:1000, GTX100443),BRN2(rabbit polyclonal, 1:1000, GTX114650) and N-cadherin (rabbit polyclonal, 1:1000, GTX127345) were purchased from Genetex.

Yin L., Ye Y., Zou L., Lin J., Dai Y., Fu Y., Liu Y., Peng Y., Gao Y., Fu Y., Qi X., Deng T., Zhang S, & Li X. (2023). AR antagonists develop drug resistance through TOMM20 autophagic degradation-promoted transformation to neuroendocrine prostate cancer. Journal of Experimental & Clinical Cancer Research : CR, 42, 204.

+ Open protocol

+ Expand

AICAR Enhances Glucose Uptake in Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Trypsin solutions, Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), antibiotic/antimycotic, and horse serum were purchased from GIBCO Life Technologies (Gaithersburg, MD, USA). We obtained 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) and Dimethyl sulfoxide (DMSO) from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). We used a Mem-PER Plus Membrane Protein Extraction Kit and 2-(N-[7-nitrobenz-2-oxa-1,3-diazol-4-yl] Amino)-2-deoxyglucose (2-NBDG) from Thermo (Sunnyvale, CA, USA). Protease inhibitor cocktail, phosphatase inhibitor cocktail, Compound C, and STO-609 were obtained from Sellbeck chemicals (Houston, TX, USA). Our siRNA was purchased from Shanghai GenePharma company (Shanghai, China). We utilized antibodies against phospho-AKT (Ser473), AMPKα (Thr172), phospho-AKT (Thr308), phospho-AS160 (Ser588), phospho-p38 MAPK (Thr180/Tyr182), phospho-ACC (Ser79), phosphoinositide 3-kinase (PI3K, P110β), AS160, AMPKα, and ACC from Cell Signaling Technology (Danvers, MA, USA); AKT, ATP1A1, p38 MAPK, and β-actin from Proteintech (Wuhan, China); and GLUT4 from Abcam (Cambridge, UK). Secondary antibodies and insulin were sourced from Yeasen Biotech (Shanghai, China). A glucose assay kit was procured from Shanghai Kexin Biotechnology Research Institute (Shanghai, China).

Li T., Lv Q., Liu C., Li C., Xie X, & Zhang W. (2024). The Lipophilic Extract from Ginkgo biloba L. Leaves Promotes Glucose Uptake and Alleviates Palmitate-Induced Insulin Resistance in C2C12 Myotubes. Molecules, 29(7), 1605.

+ Open protocol

+ Expand

In Vitro and In Vivo Compound Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols

For in vitro studies MLN0128 (Selleckchem Houston, TX, USA) was dissolved in DMSO, phenfomin was dissolved in DMEM. For in vivo studies MLN0128 was dissolved in 1-methyl-2-pyrrolidinone (NMP), then diluted in 15% PEG diluted in water; phenformin was diluted in water at 1.8 mg/ml. Rapamycin was purchased from LC Laboratories (Woburn, MA) and dissolved in DMSO. Antibodies from Cell Signaling Technology (Beverly, MA, USA) used for immunoblots were diluted 1:1,000 and included phospho-p70 S6 kinase (Thr389 #9234), phospho-S6 (Ser235/236 #4858), total 4E-BP1 (#9644), phospho-4E-BP1 (Thr37/46 #2855), phospho-Akt (Ser473 #4060), phospho-Akt (Thr308 #4056), phospho-NDRG1 (Thr346 #5482), Phospho-Tuberin/TSC2 (Thr1462 #3611), phospho-GSKα/β (Ser21/9 #9331), beta-actin (#4970), cleaved PARP (Asp214 #5625), cleaved caspase 3 (Asp175 #9661) and phospho-Raptor (Ser792 #2083). Anti-Hif-1alpha (C-Term #10006421 1:200) antibody was purchased from Cayman Chemical and anti-GLUT1 (GT11-A 1:1,000) antibody was purchased from Alpha Diagnostic International (San Antonio, TX, USA). Plasmid expressing dox-inducible 4EBP1 4Ala (pCW57.1-4EBP1_4xAla, plasmid # 38240) and control vector (pCW57.1, plasmid # 41393) were purchased from Addgene.

Momcilovic M., McMickle R., Abt E., Seki A., Simko S.A., Magyar C., Stout D.B., Fishbein M.C., Walser T.C., Dubinett S.M, & Shackelford D.B. (2015). Heightening energetic stress selectively targets LKB1-deficient non-small cell lung cancers. Cancer research, 75(22), 4910-4922.

+ Open protocol

+ Expand

Profiling Signaling Pathways in Cancer Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

H1975, HCC827, PC9 and H3255 cells were treated with serially diluted PCI32765, WZ4002 (1 μM) for 4 hours. Cells were then washed with PBS and lysed in cell lysis buffer. Phospho-EGF Receptor, Phospho-EGF Receptor (Tyr1068), Stat3, Phospho-Stat3 (Tyr705), AKT, Phospho-AKT (Ser473), Phospho-AKT (Thr308), p44/42 MAPK (Erk1/2), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204), p70 S6 Kinase, Phospho-p70 S6 Kinase (Thr389) Src, Phospho-Src Family (Tyr416), eIF4E, Phospho-eIF4E (Ser209), 4E-BP1, Phospho-4E-BP1 (Thr37/46) antibody (Cell signaling Technology) were used for immunoblotting.

Wu H., Wang A., Zhang W., Wang B., Chen C., Wang W., Hu C., Ye Z., Zhao Z., Wang L., Li X., Yu K., Liu J., Wu J., Yan X.E., Zhao P., Wang J., Wang C., Weisberg E.L., Gray N.S., Yun C.H., Liu J., Chen L, & Liu Q. (2015). Ibrutinib selectively and irreversibly targets EGFR (L858R, Del19) mutant but is moderately resistant to EGFR (T790M) mutant NSCLC Cells. Oncotarget, 6(31), 31313-31322.

+ Open protocol

+ Expand

Isolation and Characterization of Pancreatic CSCs

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

NVP-LDE225 and NVP-BEZ-235 were obtained from Chemie Tek (Indianapolis, IN). Antibodies used against Gli1, Gli2, phospho-Akt, Akt, caspase-3, PARP, E-Cadherin, N-Cadherin, Vimentin, Sox-2, Oct-4, Nanog, c-Myc, phospho-Akt, phospho-PI3K, phospho-p70S6K, phospho-GSK3Kβ, phospho-4EBP1, Akt, p70S6K, GSK3Kβ, 4EBP1, Slug, snail, snail, patched 1, patched2, Smo, Sufu Bcl-XL, Bax, Bak, Bcl2, and β-actin were purchased from Cell Signaling Technology, Inc. (Danvers, MA). Enhanced chemiluminescence (ECL) Western blot detection reagents were purchased from Thermo Fisher Scientific Corporation (Waltham, MA).
Isolation and characterization of CD44⁺CD24⁺ESA⁺ pancreatic CSCs have been described elsewhere [54 (link)]. Pancreatic CSCs were grown in well-defined stem cell culture medium [54 (link)–56 (link)].

Sharma N., Nanta R., Sharma J., Gunewardena S., Singh K.P., Shankar S, & Srivastava R.K. (2015). PI3K/AKT/mTOR and sonic hedgehog pathways cooperate together to inhibit human pancreatic cancer stem cell characteristics and tumor growth. Oncotarget, 6(31), 32039-32060.

+ Open protocol

+ Expand

Comprehensive Antibody Toolkit for Cellular Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies were purchased from Cell Signaling Technology: Akt(#4691), Phospho-Akt(Thr308)(#4056), Phospho-Akt(Ser473)(#4060), GAPDH, GSK-3β(#9315), Phospho-GSK-3β(#9323), FoxO1(#2880), Phospho-FoxO1 (Thr24)/FoxO3a(Thr32)/FoxO4(Thr28)(#2599), PRAS40(#2691), Phospho-PRAS40 (Thr246)(#2997), 4E-BP1(#9644), p70 S6K(#2708), Phospho-p70 S6K (Thr389)(#9234). SQSTM1/p62 (D5E2)(#8025), LC3B(#12513). PARP(#9532), Caspase-3(#9665), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204)(#4377), p44/42 MAPK (Erk1/2) (#4695), Phospho-NF-ΚB P65 (Ser 536)(#3033), NF-ΚB P65(#4764). Lysophosphatidic Acid (#sc-201053, Santa Cruz), Recombinant Human PDGF-BB (#220-BB, R&D), Recombinant Human C5a (Peprotech, #300-70), goat anti-IgM (#5C07615, Meridian). LC3B antibody (#3868, CST). Tubulin antibody (#8035, Santa Cruz). EEA1 antibody (#610456, BD Bioscience) and LAMP1 antibody (#24170, Abcam). Secondary antibodies, Alexa488-anti-mouse, Alexa594-anti-rabbit and Prolong gold mounting medium with DAPI were purchased from Invitrogen.

Liu X., Wang A., Liang X., Liu J., Zou F., Chen C., Zhao Z., Deng Y., Wu H., Qi Z., Wang B., Wang L., Liu F., Xu Y., Wang W., Fernandes S.M., Stone R.M., Galinsky I.A., Brown J.R., Loh T., Griffin J.D., Zhang S., Weisberg E.L., Zhang X., Liu J, & Liu Q. (2016). Simultaneous inhibition of Vps34 kinase would enhance PI3Kδ inhibitor cytotoxicity in the B-cell malignancies. Oncotarget, 7(33), 53515-53525.

+ Open protocol

+ Expand

Molecular Profiling of Glioma Therapeutics

Check if the same lab product or an alternative is used in the 5 most similar protocols

STELB was isolated from Jaspis stellifera and structurally identified it as described previously.^[^10a^] TMZ, BKM120, and ZSTK474 were purchased from Selleck Chemicals. The following antibodies were used: Caspase‐3 (Cell Signaling Technology, 9662), Cleaved Caspase‐3 (Cell Signaling Technology, 9664), PARP (Cell Signaling Technology, 9532), MGMT (Cell Signaling Technology, 86039), PTEN (Cell Signaling Technology, 9188), PI3Kα (Cell Signaling Technology, 4249), PI3Kβ (Cell Signaling Technology, 3011), PI3Kγ (Cell Signaling Technology, 5405), PI3Kδ (Cell Signaling Technology, 34050), phospho‐Akt (Thr308, Cell Signaling Technology, 13038), phospho‐Akt (Ser473, Cell Signaling Technology, 4060), phospho‐mTOR (Ser2448, Cell Signaling Technology, 5536), DNA‐PKcs (Cell Signaling Technology, 38168), Ku70 (Cell Signaling Technology, 4588), Ku80 (Cell Signaling Technology, 2180), XRCC1 (Cell Signaling Technology, 2735), γ‐H2AX (Abcam, ab81299), BRCA1 (Cell Signaling Technology, 14823, for Western blot), BRCA1 (Abcam, ab16780, for IHC), BRCA2 (Abcam, ab216972), RAD51 (Abcam, 133534), Ki‐67 (Cell Signaling Technology, 9027), HA‐Tag (Cell Signaling Technology, 3724), β‐actin (Cell Signaling Technology, 4970), anti‐rabbit IgG, HRP‐linked antibody (Cell Signaling Technology, 7074), and anti‐rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor 488 Conjugate, Cell Signaling Technology, 4412).

Peng X., Zhang S., Wang Y., Zhou Z., Yu Z., Zhong Z., Zhang L., Chen Z., Claret F.X., Elkabets M., Wang F., Sun F., Wang R., Liang H., Lin H, & Kong D. (2022). Stellettin B Sensitizes Glioblastoma to DNA‐Damaging Treatments by Suppressing PI3K‐Mediated Homologous Recombination Repair. Advanced Science, 10(3), 2205529.

+ Open protocol

+ Expand

Characterization of Cellular Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

MK2206, zaragozic acid, and myriocin were obtained from Cayman Chemical. Oil Red O and Alizarin Red stainings were bought from Sigma-Aldrich. The Alkaline Phosphatase Assay Kit was purchased from Abcam. Antibodies used were as follows: caveolin (polyclonal; Santa Cruz Biotechnology), Na⁺,K⁺ adenosine triphosphatase (polyclonal; Cell Signaling), calnexin (polyclonal; Abcam), GM130 (polyclonal; Cell Signaling), LAMP1 (polyclonal; Abcam), RUNX2 (polyclonal; Cell Signaling), GOLGA7 (monoclonal; Abcam), Sec61a (Abcam), hexokinase II (polyclonal; Cell Signaling), phospho-Akt (pS473; polyclonal; Cell Signaling), phospho-Akt (pS473; monoclonal; Cell Signaling), and Akt (polyclonal; Cell Signaling). The plasmids for Lyn-AktAR and Kras-AktAR were provided by J. Zhang (University of California, San Diego). The plasmid for GFP-PH-Akt was a gift from T. Balla [National Institutes of Health (NIH)].

Levental K.R., Surma M.A., Skinkle A.D., Lorent J.H., Zhou Y., Klose C., Chang J.T., Hancock J.F, & Levental I. (2017). ω-3 polyunsaturated fatty acids direct differentiation of the membrane phenotype in mesenchymal stem cells to potentiate osteogenesis. Science Advances, 3(11), eaao1193.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!