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Nvp aew541

Manufactured by Cayman Chemical
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NVP-AEW541 is a chemical compound used in research applications. It is a selective inhibitor of the PI3Kα isoform. NVP-AEW541 is supplied as a crystalline solid.

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Lab products found in correlation

Linsitinib, by LC Laboratories (2 mentions) Ly3009120, by Selleck Chemicals (1 mentions) Azd6244, by Selleck Chemicals (1 mentions) Picropodophyllotoxin, by Merck Group (1 mentions) Igf 1, by Merck Group (1 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) Tamoxifen, by Merck Group (1 mentions) Rapamycin, by Merck Group (1 mentions) Pmirtarget vector, by OriGene (1 mentions) Fibronectin, by Merck Group (1 mentions) Akt1 2 inhibitor, by Merck Group (1 mentions) Actinomycin d, by Merck Group (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) U0126, by Merck Group (1 mentions) Mk 2206, by Cayman Chemical (1 mentions) Gf109203x, by Cayman Chemical (1 mentions) Protein g sepharose bead, by GE Healthcare (1 mentions) Hrp conjugated and fluorophore conjugated secondary antibodies, by Jackson ImmunoResearch (1 mentions) Glosensor, by Promega (1 mentions) Ci 1040, by Cayman Chemical (1 mentions)

9 protocols using nvp aew541

1

Synergistic Combinatorial Drug Screening

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For the synergy assay, cells seeded the day before were treated with different concentrations of AZD6244 (0.1, 1, 10, 30 and 50 μM, Selleck Chemicals) in combination with NVP-AEW541 (0.1, 0.3, 1, 3 and 10 μM, Cayman Chemical) or LY3009120 (0.1, 0.3, 1, 3 and 15 μM, Selleck Chemicals). The synergy scores were determined using the R package synergyfinder [56 (link)] with the relative growth rates thresholded between 0 and 1 as input (0 meaning no growth or cell death and 1 meaning growth as fast as the DMSO control).
Dorel M., Klinger B., Mari T., Toedling J., Blanc E., Messerschmidt C., Nadler-Holly M., Ziehm M., Sieber A., Hertwig F., Beule D., Eggert A., Schulte J.H., Selbach M, & Blüthgen N. (2021). Neuroblastoma signalling models unveil combination therapies targeting feedback-mediated resistance. PLoS Computational Biology, 17(11), e1009515.
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2

Investigating Growth Factors and Inhibitors

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Insulin-like growth factor 1 (IGF-1) (catalog I3769), picropodophyllotoxin (catalog T9576), ATP (catalog A1862), and tamoxifen (catalog T5648) were purchased from (Sigma-Aldrich Chemicals, Sigma-Aldrich, St. Louis, MO, USA). NVP-AEW 541 (catalog 13641) was purchased from Cayman chemicals. Radiolabeled glutamate (catalog NET490250UC) was purchased from Perkin Elmer, while unlabeled glutamatic acid (catalog A1222A) was purchased from ThermoFisher Scientific.
Prabhu D., Khan S.M., Blackburn K., Marshall J.P, & Ashpole N.M. (2019). Loss of insulin-like growth factor-1 signaling in astrocytes disrupts glutamate handling. Journal of neurochemistry, 151(6), 689-702.
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3

Investigating IGF-I/II Signaling Pathways

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IGF-I and IGF-II were from Preprotech (Rocky Hill, NJ), lipofectamine 2000 and lipofectamine RNAiMax from Life Technologies Inc. Laboratories (Paisley, UK), scramble (miR-NC), synthetic pre-miR-199a-5p (miR-199a-5p), and mirVana miR-199a-5p inhibitor were purchased from Ambion (Applied Biosystems, CA, USA), fibronectin from Sigma-Aldrich (Saint Louis Missouri, USA).
Constructs encoding either pcDNA.3.1-HA-myr-AKT dominant active construct (AKT) or the empty vector pcDNA3.1 (vector) were kindly provided by prof. P. Tassone (University “Magna Graecia” of Catanzaro).
Construct encoding 3′UTR clone of DDR1 in pMirTarget Vector, the related empty vector (pCMV6) and the human IGF-II cDNAs were from OriGene Technologies (Rockville, USA). The following kinase inhibitors were used: the IGF-IR inhibitor NVP-AEW541 (Cayman Chemical, Ann Arbor, USA); the PI3 kinase inhibitor LY 294002 (Calbiochem, Merck Millipore, Nottingham, UK); the MEK1 inhibitor U0126 (Sigma-Aldrich, Saint Louis Missouri, USA); the TORC1 inhibitor rapamycin (Sigma-Aldrich); the AKT 1,2 inhibitor (Sigma-Aldrich, Saint Louis Missouri, USA). Actinomycin D and cycloheximide were purchased from Sigma-Aldrich, MTT from Amersham Bioscences.
Matà R., Palladino C., Nicolosi M.L., Presti A.R., Malaguarnera R., Ragusa M., Sciortino D., Morrione A., Maggiolini M., Vella V, & Belfiore A. (2015). IGF-I induces upregulation of DDR1 collagen receptor in breast cancer cells by suppressing MIR-199a-5p through the PI3K/AKT pathway. Oncotarget, 7(7), 7683-7700.
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4

Ubiquitination Pathway Regulation in Cells

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The primers were synthesized and PAGE purified by IDT DNA. Fugene 6 transfection reagent was purchased from Promega while Lipofectamine RNAiMax was from Life Technologies/Invitrogen. All cell culture reagents including media, antibiotics were from GIBCO/Invitrogen. Antibodies to total Ubiquitin (clone P4D1) was from Santacruz Biotechnology, Luciferase (Millipore), pAKT, AKT (Cell Signaling), Ubiquitin, Lys63 specific (clone Apu3, Millipore), His-tag (clone H3 and C-term, Invitrogen) or Millipore (H8 clone). FLAG tag-HRP antibody (Clone M2) was purchased from Sigma Aldrich. HRP-conjugated and fluorophore-conjugated secondary antibodies were from Jackson ImmunoResearch. ON-TARGET plus siRNA to TRAF6 and scrambled control siRNA were obtained from GE Life Sciences/Dharmacon. D-Luciferin was from Xenogen Corp while GloSensor was from Promega. Erlotinib was a kind gift from Genentech. Tyrphostin AG1478 was obtained from Cayman Chemical. TNF-α antagonist (WP9QY) and IL-1R antagonist were from SantaCruz biotechnology. Recombinant IGF-1, IL-1α were from PepreoTech. CI-1040, GF109203X, NVP-AEW541 and MK2206 (Cayman Chemical), and Linsitinib (LC Laboratories). Protein A and Protein G sepharose beads were from GE HealthCare. Purified NEDD4-1 (Sigma Aldrich), Myc-tagged ubiquitin (Cat. No. U-115) UBE1 (Cat. No. E305), and UBCH5 (Cat. No. E2-616) all were from Boston Biochemicals.
Nyati S., Chaudhry N., Chatur A., Gregg B.S., Kimmel L., Khare D., Basrur V., Ray D, & Rehemtulla A. (2018). A novel reporter for real-time, quantitative imaging of AKT-directed K63-poly-ubiquitination in living cells. Oncotarget, 9(13), 11083-11099.
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5

Inhibition of Signaling Pathways in Zebrafish

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To inhibit the endpoint of the Wnt signaling pathway, ectopic expression of the dominant negative TCF from the transgenic Tg(HS:TCFΔC)W74 (Martin and Kimelman, 2012 (link)) was induced by increasing temperature to 37° C for 20 minutes, followed by return to 28.5° C embryo media (embryos) or system water (one week post-embryonic fish). WT controls were heat shocked for the same period in as the DN TCF.
EGF signaling was inhibited for 2 hour periods with the cell-permeable EGF kinase inhibitor AG-1478 (Cayman Chemical 10010244) at the previously reported concentration of 3μM solubilized in DMSO (Budi et al., 2008 ). Control embryos were exposed to an equal concentration of DMSO for the same 2-hour periods. Relative expression of myelin basic protein between control and AG-1478 inhibited embryos was compared for internal control to demonstrate activity of EGF inhibition.
IGF signaling was inhibited for 2 hour periods with NVP-AEW541 (Cayman Chemical 13641) at the previously reported concentration of 5 mM solubilized in DMSO (Chablais and Jazwinska, 2010 ). Control embryos were exposed to an equal concentration of DMSO for the same 2-hour periods. Intensity of phosphorylated IGF 1 receptor was compared between control and NVP-AEW541 inhibited embryos for an internal control to demonstrate activity of IGF inhibition.
Li J., Dedloff M.R., Stevens K., Maney L., Prochaska M., Hongay C.F, & Wallace K.N. (2019). A novel group of secretory cells regulates development of the immature intestinal stem cell niche through repression of the main signaling pathways driving proliferation.. Developmental biology, 456(1), 47-62.
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6

IGF-1 Stimulation and PI3K Inhibition

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Cells were stimulated for 20 min with recombinant human IGF-1 (R&D systems) at a final concentration of 100 ng/ml. PI3K inhibitors BKM-120 (Selleck Chemicals), BYL-719 (Selleck Chemicals), A66 (Selleck Chemicals), TGX-221 and IC87114 [gifts from Peter Shepherd, University of Auckland, New Zealand; (54 (link))] were added to cells 15 min prior to IGF-1 stimulation at final concentration of 1 μM. AKT inhibitor MK-2206 (Active Biochem) was added to cells 15 min prior to IGF-1 stimulation at final concentration of 1 μM. NVP-AEW541 (Cayman Chemical) and GSK2334470 (Tocris) were added to cells 15 min prior to growth factor stimulation at final concentration of 1 μM.
Clement E., Inuzuka H., Nihira N.T., Wei W, & Toker A. (2018). Skp2-dependent reactivation of AKT drives resistance to PI3K inhibitors. Science signaling, 11(521), eaao3810.
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7

Inhibiting IGF-1 Signaling and Assessing Cell Viability

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Cells were treated with NVP-AEW541 (Cayman Chemical) and OSI-906 (ChemieTek) at the indicated concentrations. Prior to treatment with IGF-1, cells were kept in serum-free media for two hours in combination with the IGF-1 inhibitor. Cells were then treated with IGF-1 (Cell Signaling Technologies) for 15 min and lysed in RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 0.1% SDS) supplemented with 200 mM NaVO4 and 50 mM NaF. Cell extracts were separated by SDS-PAGE and blotted with anti-phospho AKT and imaged (LiCor). For assessment of cell viability, EWS502 cells were transduced with lentiviral pLL5.0-PTEN or pLL5.0 as a vector control. 24 hours post infection the cells were treated with NVP-AEW541 in complete media. Viability was assayed 72 hours following NVP-AEW541 treatment using WST-1 (Roche).
Patel M., Gomez N.C., McFadden A.W., Moats-Staats B.M., Wu S., Rojas A., Sapp T., Simon J.M., Smith S.V., Kaiser-Rogers K, & Davis I.J. (2014). PTEN deficiency mediates a reciprocal response to IGF-1 and mTOR inhibition. Molecular cancer research : MCR, 12(11), 1610-1620.
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8

Combination Therapy Optimization

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Buparlisib (BKM120) was provided by MTA from Novartis Pharma (Basel, Switzerland) or obtained from Selleck Chemical. Doxorubicin HCl was obtained from Shandong Tianyu Fine Chemical Co., Ltd. NVP-AEW541 was obtained from Cayman Chemical. Rapamycin was obtained from Cell Signaling Technology. Trametinib and pictilisib (GDC-0941) were obtained from Selleck Chemical.
Anderson J.L., Park A., Akiyama R., Tap W.D., Denny C.T, & Federman N. (2015). Evaluation of In Vitro Activity of the Class I PI3K Inhibitor Buparlisib (BKM120) in Pediatric Bone and Soft Tissue Sarcomas. PLoS ONE, 10(9), e0133610.
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9

Egg Yolk-based Compound Screening Protocol

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For egg yolk feeding, chicken eggs were obtained from local grocery stores, and the yolk was separated and diluted to 5% by volume with 0.3× Danieau solution as previously described (26 (link)). All drugs were made in 1,000× stock solution and stored in light-protected Eppendorf tubes at −20°C. NBI-31772 (5 mmol/L; Sigma-Aldrich), linsitinib (10 mmol/L; LC laboratories), H-89 (10 mmol/L; LC laboratories), AICAR (100 mmol/L; LC laboratories), afatinib (10 mmol/L; LC laboratories), SB431542 (10 mmol/L; Selleckchem), DAPT (10 mmol/L; Sigma-Aldrich), SU5402 (15 mmol/L; Calbiochem and Tocris), neocuproine (10 mmol/L; Sigma-Aldrich), PD0325901 (10 mmol/L; Sigma-Aldrich), U0126 (10 mmol/L; LC laboratories), and TUDCA (0.5 mol/L; Calbiochem) were dissolved in DMSO at the indicated concentrations. NVP-AEW541 (10 mmol/L; Cayman Chemicals), vatalanib (10 mmol/L; LC Laboratories), and SAG (10 mmol/L; EMD Millipore) were dissolved in water. CyA (10 mmol/L; LC Laboratories) was dissolved in ethanol.
Induction of transgene expression of Kir6.2DN was performed as previously described (24 (link)). SU5402 was added after 16 h of the induction for an 8-h treatment.
Li M., Page-McCaw P, & Chen W. (2015). FGF1 Mediates Overnutrition-Induced Compensatory β-Cell Differentiation. Diabetes, 65(1), 96-109.
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