Rad001

Manufactured by Selleck Chemicals

Sourced in United States, China

RAD001 is a laboratory equipment used for analyzing and measuring various chemical and biological samples. It is designed to provide accurate and precise measurements of specific parameters within the samples. The core function of RAD001 is to perform analytical tasks, but a more detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Close spelling variants of this product

Everolimus (RAD001)

46 protocols using rad001

NSCLC Cell Line Characterization and Drug Testing

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The H23, H1703, A549, HCC827, H157, H460, H2228, H3122 and H322 NSCLC cell lines were obtained from American Type Tissue Collection (Manassas, VA), and were grown in RPMI 1640 supplemented with 10% FBS and 1× Antibiotic/Antimycotic (Invitrogen, Carlsbad, CA). Among them, Cell lines H23, HCC827, H1703 and A549 were tested and authenticated by short tandem repeat profiling in August, 2014 and 2015 and the rest were not authenticated. PC9 cells were a gift from Dr. Susumu Kobayashi, Harvard Medical School, Boston, MA. Ba/F3 cells were obtained from G. Gilliland (Brigham and Women's Hospital, Boston, Massachusetts, USA). RICTOR-dependent Ba/F3 cells were developed by transfecting Ba/F3 cells with a doxycycline-inducible plasmid for the overexpression of wild-type RICTOR in the presence of doxycycline (RICTOR-pTetOne, modified from Tet-One Systems, Clontech Laboratories, Inc, CA, US and generated by GENEWIZ, Inc, NJ, US). BEZ235, AZD2014, RAD001 (everolimus), erlotinib, MK2206 and LY294002 were obtained from Selleck Chemicals (Houston, TX); Cisplatin was obtained from Sigma-Aldrich (St. Louis, MO).

Cheng H., Zou Y., Ross J.S., Wang K., Liu X., Halmos B., Ali S.M., Liu H., Verma A., Montagna C., Chachoua A., Goel S., Schwartz E.L., Zhu C., Shan J., Yu Y., Gritsman K., Yelensky R., Lipson D., Otto G., Hawryluk M., Stephens P.J., Miller V.A., Piperdi B, & Perez-Soler R. (2015). RICTOR amplification defines a novel subset of lung cancer patients who may benefit from treatment with mTOR1/2 inhibitors. Cancer discovery, 5(12), 1262-1270.

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Combinatorial Drug Screening Protocol

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AZD1775 and LY2606368 were manufactured by MD Anderson’s Institute for Applied Chemical Science. TP0903 was provided by Tolero Pharmaceuticals. Temozolomide, cabozantinib, and RAD001 were obtained from Selleck Chemicals. All compounds were dissolved in dimethyl sulfoxide for in vitro treatments.

Sen T., Tong P., Diao L., Li L., Fan Y., Hoff J., Heymach J.V., Wang J, & Byers L.A. (2017). Targeting AXL and mTOR Pathway Overcomes Primary and Acquired Resistance to WEE1 Inhibition in Small-Cell Lung Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(20), 6239-6253.

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NMDA Receptor Pharmacology Assay

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N-methyl-D-aspartate (NMDA) was purchased from Sigma-Aldrich (St. Louis, MO). RAD001 and MK2206 were purchased from Selleckchem (Houston, TX). AS1842856 was purchased from EMD Millipore (Billerica, MA). LiCl and dimethyl sulfoxide (DMSO) were purchased from Thermo Fisher Scientific (Waltham, MA).

Swiatkowski P., Nikolaeva I., Kumar G., Zucco A., Akum B.F., Patel M.V., D’Arcangelo G, & Firestein B.L. (2017). Role of Akt-independent mTORC1 and GSK3β signaling in sublethal NMDA-induced injury and the recovery of neuronal electrophysiology and survival. Scientific Reports, 7, 1539.

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Chemical Inhibitors for Cell Signaling

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LY294002 was obtained from Cell Signaling Technology (Danvers, MA). RAD001, MK-2206 and VPS34-IN1 were obtained from Selleck (Shanghai, China).

Wang H., Huang F., Zhang Z., Wang P., Luo Y., Li H., Li N., Wang J., Zhou J., Wang Y, & Li S. (2019). Feedback Activation of SGK3 and AKT Contributes to Rapamycin Resistance by Reactivating mTORC1/4EBP1 Axis via TSC2 in Breast Cancer. International Journal of Biological Sciences, 15(5), 929-941.

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Knockdown of CCKBR in A431 Cells

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The human epidermoid carcinoma A431 cell line, which overexpresses CCKBR, was generated and kindly provided by Dr. Luigi Aloj 23 (link). The A431 and A431/CCKBR cell lines were cultured in DMEM, whereas the rat pancreatic acinar AR42J cells (ECACC, UK) were grown in RPMI medium, supplemented with 10% FCS, 2 mM glutamine and antibiotics (0.1 mg/mL streptomycin, 100 IU penicillin) at 37 °C and 5% CO₂. For CCKBR-specific knock-down, duplex siRNAs against CCKBR or control duplex against luciferase (Microsynth) were used at a final concentration of 100 nM in Optimem (Gibco): CCKBRseq1 sense RNA 5′-UAUACGAGUAGUAGCACCAdTdT-3′, CCKBRseq2 sense RNA 5′-CCGCCAAAGGAUGGAGUACdTdT-3′ and control sense RNA 5′-CGUACGCGGAAUACUUCGAdTdT-3′. Cells at 60-80% confluence were transfected with siRNAs by using Lipofectamin 3000 according to the manufacturer's recommendations. RAD001 (Selleckchem), rapamycin (Enzo Life Sciences), BML-257(Enzo), SC-514 (Enzo) or metformin (Selleckchem) were diluted in DMSO or water, respectively.

Grzmil M., Qin Y., Schleuniger C., Frank S., Imobersteg S., Blanc A., Spillmann M., Berger P., Schibli R, & Behe M. (2020). Pharmacological inhibition of mTORC1 increases CCKBR-specific tumor uptake of radiolabeled minigastrin analogue [177Lu]Lu-PP-F11N. Theranostics, 10(24), 10861-10873.

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Modulating IGF-1R/InsR Signaling in Breast Cancer

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Parental cell lines were obtained from ATCC and cultured in DMEM (Gibco) with 10% FBS (Hyclone) for <3 months before analysis. Long-term estrogen-deprived (LTED) cells were generated through culture in phenol red-free DMEM with 10% dextran-charcoal-treated FBS (DCC-FBS) for 3 months. Cells resistant to the anti-estrogen fulvestrant (fulv; MCF-7/FR, T47D/FR) were gifts from Matthew Ellis (Washington University), and maintained in DMEM/10% FBS with 1 μM fulv. LTED and FR cells were authenticated by STR genotyping (Univ. of Vermont Advanced Genome Technologies Core). Cells were transfected with siRNA targeting IGF-1R (#GS3480), InsR (#GS3643), IRS-1 (#GS3667), IRS-2 (#GS8660), or non-silencing control (#1027310) from Qiagen using Lipofectamine RNAiMAX per manufacturer’s instructions (Life Technologies). Cells were treated with phenol red-free DMEM containing 10% dextran-charcoal-treated FBS (DCC-FBS; Hyclone), RAD001, OSI-906 (Selleck Chemicals), fulv (Tocris Bioscience), IGF-1 (R&D Systems), or 17β-estradiol (E2; Sigma) as indicated.

Yang W., Schwartz G.N., Marotti J.D., Chen V., Traphagen N.A., Gui J, & Miller T.W. (2018). Estrogen receptor alpha drives mTORC1 inhibitor-induced feedback activation of PI3K/AKT in ER+ breast cancer. Oncotarget, 9(10), 8810-8822.

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Characterization of Apoptosis-Inducing Agents

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Imatinib, nilotinib (AMN107), and BEZ235 were kindly provided by Dr. E. Buchdunger and Dr. P. Manley (Novartis Pharma AG, Basel, Switzerland). Ponatinib and RAD001 (everolimus) were purchased from SelleckChem (Houston, TX, USA) and GX015-070 (obatoclax) from ChemieTek (Indianapolis, IN, USA). Stock solutions of drugs were prepared by dissolving in dimethyl-sulfoxid from Sigma-Aldrich (St. Louis, MO, USA). RPMI 1640 medium was purchased from Lonza (Verviers, Belgium), fetal bovine serum (FBS) from Thermo Fisher Scientific (Waltham, MA, USA), and bovine serum albumin (BSA) from Sigma-Aldrich. Propidium Iodide (PI) was purchased from Sigma-Aldrich, Annexin V from Affymetrix (Santa Clara, CA, USA), and ³H-thymidine from Perkin Elmer (Waltham, MA, USA). A characterization of polyclonal and monoclonal antibodies (mAb) used in this study is provided in Supplementary Table 5.

Stefanzl G., Berger D., Cerny-Reiterer S., Blatt K., Eisenwort G., Sperr W.R., Hoermann G., Lind K., Hauswirth A.W., Bettelheim P., Sill H., Melo J.V., Jäger U, & Valent P. (2017). The pan-BCL-2-blocker obatoclax (GX15-070) and the PI3-kinase/mTOR-inhibitor BEZ235 produce cooperative growth-inhibitory effects in ALL cells. Oncotarget, 8(40), 67709-67722.

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Cell Viability Assay for Leukemia Drugs

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Cells (25,000 per well) were seeded in 100 ml of complete growth medium in 96-well plates and incubated with chemotherapeutic agents or vehicle. T-ALL cells were split every 48 hr and AML cells were split every 72 hr. Cell viability was assessed by counting viable cells based on trypan blue vital dye staining (Invitrogen), according to the manufacturer’s instructions. Chemotherapeutic drugs included: asparaginase (pegaspargase, Shire, Lexington, MA), dexamethasone (Sigma-Aldrich), vincristine (Selleckchem, Houston, TX), doxorubicin (Sigma-Aldrich), 6-mercaptopurine (Abcam, Cambridge, UK), CHIR99021 (Selleckchem), rapamycin (Selleckchem), RAD001 (Selleckchem), AZD2014 (Selleckchem), thapsigargin (Sigma-Aldrich) and Wnt3A (R&D systems, Minneapolis, MN). BRD0705 and BRD3731 were synthesized as described (Wagner et al., 2018 ). Caspase 3/7 activity was assessed using the Caspase Glo 3/7 Assay (Promega, Madison, WI) according to the manufacturer’s instructions.

Hinze L., Pfirrmann M., Karim S., Degar J., McGuckin C., Vinjamur D., Sacher J., Stevenson K.E., Neuberg D.S., Orellana E., Stanulla M., Gregory R.I., Bauer D.E., Wagner F.F., Stegmaier K, & Gutierrez A. (2019). Synthetic Lethality of Wnt Pathway Activation and Asparaginase in Drug-Resistant Acute Leukemias. Cancer cell, 35(4), 664-676.e7.

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Comprehensive Signaling Pathway Analysis

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Antibodies against AKT1, AKT2, AKT3, panAKT, pAKT S473, pAKT T308, pAKT1 S473, pAKT2 S474, pS6 S240/244, pGSK3α/β S21/9, pHER2 Y877, S100A4, MMP2, RANK, and CTGF, and secondary HRP-linked antibodies against rabbit or mouse IgG were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Antibody against AKT3 was provided by Millipore (Burlington, MA, USA). Antibody against HSC-70 was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Antibody against pDDR1/2 Y796/Y740 was provided by R&D systems Inc. (Minneapolis, MN, USA). RAD001 was provided by Selleck Chemicals (Houston, TX, USA) and MK-2206 was obtained from AbMole BioScience Inc. (Houston, TX, USA). Recombinant human TGF-β1 and recombinant human EGF were purchased from PeproTech Inc. (Rocky Hill, NJ, USA).

Hinz N., Baranowsky A., Horn M., Kriegs M., Sibbertsen F., Smit D.J., Clezardin P., Lange T., Schinke T, & Jücker M. (2021). Knockdown of AKT3 Activates HER2 and DDR Kinases in Bone-Seeking Breast Cancer Cells, Promotes Metastasis In Vivo and Attenuates the TGFβ/CTGF Axis. Cells, 10(2), 430.

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Synthesis and Characterization of Multifunctional Nanocarriers

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RAD001 was purchased from Selleck Chemicals (Houston, TX). Trimethylamine (Et₃N), 1,4-butanediamine, cystamine dihydrochloride, dithiothreitol (DTT), hydroxyl N-hydroxylsuccinimide polyethylene glycol (NHS-PEG-OH, molecular weight [MW] = 2000 Da), eight-armed PEG (bPEG, MW = 15 000 Da), 1,1′-carbonyldiimidazole (CDI), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (Saint Louis, MO). Third-generation polyamidoamine dendrimers (G3 PAMAM) was purchased from Dendritech (Midland, MI). 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) was obtained from Beyotime Biotechnology (Haimen, China). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl), 4-(dimethylamino)-pyridine (DMAP), and fluorescein isothiocyanate (FITC) were purchased from Aladdin Industrial (Shanghai, China). Dialysis membranes were obtained from Spectrum Laboratories (Rancho Dominguez, CA). Unmodified and fluorescence-tagged negative-control small interfering RNA (nc-siRNA, nc-siRNA^Cy5.5+, and nc-siRNA^FITC+) was purchased from Biomics Biotechnologies (Nantong, China).

Yuan Y., Jin P., Wang Y., Zhao X., Hu Q., Wu W., Huang J, & Zhang N. (2020). A dendritic, redox-responsive, supramolecular (Dr.S) system for lysis-triggered delivery for drug-resistant renal cancer. RSC Advances, 10(62), 37826-37833.

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