Cycloheximide was purchased from Sigma-Aldrich (St. Louis, MO). The mitogen-activated protein kinase (MAPK)/ERK kinase (MEK) inhibitor, PD98059, was purchased from Calbiochem (La Jolla, CA). The PI3K inhibitor, LY294002, was purchased from Cayman Chemical (Ann Arbor, MI). The AKT inhibitor, Perifosine (>99 % pure), was purchased from LC laboratories (Woburn, MA). PTEN phosphatase inhibitor SF1670 was purchased from Cayman Chemical (Ann Arbor, MI). Nerve growth factor (NGF 2.5S) was purchased from Life Technologies (Grand Island, NY).
Perifosine
Manufactured by LC Laboratories
Sourced in United States
Perifosine is a synthetic organic compound that functions as a protein kinase B (Akt) inhibitor. It is a pharmaceutical ingredient used in research and development applications.
Automatically generated - may contain errors
Lab products found in correlation
Nerve growth factor ngf 2.5s, by Thermo Fisher Scientific (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Ly294002, by Cayman Chemical (1 mentions)
Wortmannin, by Merck Group (1 mentions)
Bkm120, by Chemietek (1 mentions)
Everolimus, by Merck Group (1 mentions)
Nvp bez235, by LC Laboratories (1 mentions)
Gdc 0941, by LC Laboratories (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Skbr3, by American Type Culture Collection (1 mentions)
Wnt3a, by R&D Systems (1 mentions)
Hcc1937, by American Type Culture Collection (1 mentions)
Caspase 8, by Cell Signaling Technology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Z vad fmk, by Apexbio (1 mentions)
N acetyl cysteine (nac), by Cayman Chemical (1 mentions)
Caspase 9, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Anti parp, by Cell Signaling Technology (1 mentions)
4 protocols using perifosine
1
Analysis of MAPK/PI3K Signaling Inhibitors
2
Preparation of Perifosine and Trametinib
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Perifosine was purchased from LC Laboratories (Woburn, MA), and trametinib was provided by the Developmental Therapeutics Program at the National Cancer Institute (Bethesda, MD). Stock concentrations of Perifosine were prepared in Dulbecco’s modified Eagle’s medium at 10 mM and diluted to working concentrations in tumor stem media. Stock trametinib was prepared in DMSO at 10 mM and diluted to working concentrations in tumor stem media. Stock concentrations were stored in aliquots at −20°C.
3
Breast Cancer Cell Lines Cultivation
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The breast cancer cell lines MDA-MB-231 (triple negative), HCC1937 (triple negative), MCF-7 (ER-positive) and SK-BR3 (HER2-positive) were purchased from American Type Culture Collection (ATCC) (Manassas, VA). MDA-MB-231 and MCF-7 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM), HCC1937 cells were cultured in RPMI140 and SK-BR3 cells were cultured in MaCoy's 5A with 10% fetal bovine serum and antibiotics. The cells were incubated at 37°C under 5% CO2. The compounds used in this study were purchased from the following vendors: Wortmannin (Sigma-Aldrich, St. Louis, MO), GDC-0941 (LC Laboratories, Woburn, MA), Perifosine (LC Laboratories, Woburn, MA), Everolimus (Sigma-Aldrich, St. Louis, MO), NVP-BEZ235 (LC Laboratories, Woburn, MA), BKM120 (Chemie Tek, Indianapolis, IN), XL147 (Chemie Tek, Indianapolis, IN), LGK974 (StemRd, Burlingame, CA), and WNT3A (R&D Systems, Minneapolis, MN).
4
Apoptosis Induction by Phospholipid
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Cells were treated with DMSO or PL for various durations. The process for determining apoptosis with flow cytometry was referred to as previously reported [52 (link)]. The protein expressions involved in the mechanisms that PL-mediated apoptosis were examined with Western blots. caspase-3, caspase-8, caspase-9, anti-PARP, and anti-GAPDH antibodies were purchased from Cell Signaling Inc. (Danvers, MA, USA). GAPDH was used as a loading control. Z-VAD-FMK, a pan-caspase inhibitor, was purchased from ApexBio Technology LLC (Houston, TX, USA). The inhibitor of ROS, NAC, was purchased from Cayman. Moreover, the MAPK and Akt signaling pathways, including the total forms and phosphorylated forms of ERK, JNK, p38, Akt, and mTOR (all antibodies were purchased from Cell Signaling), were examined in the cells after PL treatment. PD98058 and perifosine, inhibitors of Erk and Akt, respectively, were purchased from LC Laboratories (Woburn, MA, USA). The plasmid, including pBSSK+ and constitutively active human Akt1 construct (pmAkT), were used as previously reported [50 (link)]. Three independent assays were performed.
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