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Vx 11e

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The VX-11e is a laboratory centrifuge designed for general-purpose applications. It can separate different components of a liquid mixture based on their density differences. The centrifuge has a maximum speed of 6,000 RPM and can accommodate sample volumes up to 100 mL. It is equipped with a digital display and various safety features.

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Lab products found in correlation

Fr180204, by Bio-Techne (2 mentions) Sch772984, by Selleck Chemicals (2 mentions) Plx4720, by Selleck Chemicals (1 mentions) Gsk1120212, by Chemietek (1 mentions) Azd6244, by Chemietek (1 mentions) Paclitaxel, by Thermo Fisher Scientific (1 mentions) Nitrotetrazolium blue chloride, by Merck Group (1 mentions) Bacto agar, by BD (1 mentions) Mk 2206, by Chemietek (1 mentions) Pd0325901, by Chemietek (1 mentions) Q vd oph, by R&D Systems (1 mentions) Gdc 0941, by Chemietek (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Dabrafenib, by Selleck Chemicals (1 mentions) Selumetinib, by Selleck Chemicals (1 mentions) Trametinib, by Selleck Chemicals (1 mentions) Vemurafenib, by Selleck Chemicals (1 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions)

7 protocols using vx 11e

1

Cell Culture and Compound Treatment

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A375, 293T, SKMEL-19, and WM266.4 cells were grown in DMEM with 10% FBS. A375 and 293T cells were acquired from the ATCC. WM266.4 cells were acquired through the Cancer Cell Line Encyclopedia (20 ). SKMEL-19 cells were a gift from N. Rosen (Memorial Sloan Kettering Cancer Center, New York, NY). GSK1120212, GSK2118436, VX-11e, and AZD6244 were obtained from Chemietek. PLX4720 was obtained from Selleck. SCH772984 was synthesized by J & W PharmLab. Lentiviral production and infection were performed as previously described (19 (link)).
Goetz E.M., Ghandi M., Treacy D.J., Wagle N, & Garraway L.A. (2014). ERK Mutations Confer Resistance to Mitogen-Activated Protein Kinase Pathway Inhibitors. Cancer research, 74(23), 7079-7089.
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2

Anchorage-Independent Cancer Cell Proliferation

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Anchorage-independent proliferation was quantified by soft agar assay. Cells were suspended in 1 mL of 0.36 % (w/v) bacto-agar (BD Biosciences) overlaid by a 0.6 % (w/v) bacto-agar support in 6-well plates in triplicates at densities of 5 × 103 cells per well for A375, A375.S2, A375Scr, A375 shFN2, A375.S2 Scr, A375.S2 shFN2 lines, respectively. Plates were incubated at 37 °C in normoxic conditions. Pathway inhibitors were used at the following concentrations: 3.0 μM cisplatin (DNA-crosslinking agent), 3.0 μM VX-11e (ERK inhibitor), 3.0 μM vemurafenib (BRAFV600E inhibitor) (Chemietek, Indianapolis, IN), and 50 nM paclitaxel (microtubule stabilizing agent) (Life Technologies). Pathway inhibitors were administered at the beginning of the experiment or initiated 14 days later. At the end of the experiment, wells were stained with 0.5 μg/mL of nitrotetrazolium blue chloride (Sigma-Aldrich). Colonies with > 20 μm in diameter were counted using the Bioreader Software and a BioSys BioCount 4000 Pro machine. Clonogenic survival for each line and condition was plotted as frequency of colony size normalized to a gaussian distribution. Experiments were performed in triplicate.
Afasizheva A., Devine A., Tillman H., Fung K.L., Vieira W.D., Blehm B.H., Kotobuki Y., Busby B., Chen E.I, & Tanner K. (2016). Mitogen-activated protein kinase signaling causes malignant melanoma cells to differentially alter extracellular matrix biosynthesis to promote cell survival. BMC Cancer, 16, 186.
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3

Chemical Compounds Utilized in Research

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SCH772984 was purchased from Selleckchem (Cat. No. S7101). VX11e was purchased from ChemieTek (Cat. No. CT-VX11e). FR180204 was purchased from Tocris (Cat. No 3706).
Chaikuad A., Tacconi E., Zimmer J., Liang Y., Gray N.S., Tarsounas M, & Knapp S. (2014). A unique inhibitor binding site in ERK1/2 is associated with slow binding kinetics. Nature chemical biology, 10(10), 853-860.
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4

Inhibitors of Cell Signaling Pathways

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Romidepsin was obtained from the Developmental Therapeutics Program of the National Cancer Institute (Bethesda MD). The MEK inhibitor PD-0325901, the AKT inhibitor MK-2206, the ERK inhibitor Vx-11e and the PI3K inhibitor GDC-0941 were purchased from ChemieTek (Indianapolis, IN). Q-VD-OPh was from R&D Systems (Minneapolis, MN). The dual pathway inhibitor, D-87503, was synthesized in-house by the Chemical Biology Laboratory, Center for Cancer Research (Frederick, MD).
Bahr J.C., Robey R.W., Luchenko V., Basseville A., Chakraborty A.R., Kozlowski H., Pauly G.T., Patel P., Schneider J.P., Gottesman M.M, & Bates S.E. (2016). Blocking downstream signaling pathways in the context of HDAC inhibition promotes apoptosis preferentially in cells harboring mutant Ras. Oncotarget, 7(43), 69804-69815.
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5

Chemical Compounds Utilized in Research

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SCH772984 was purchased from Selleckchem (Cat. No. S7101). VX11e was purchased from ChemieTek (Cat. No. CT-VX11e). FR180204 was purchased from Tocris (Cat. No 3706).
Chaikuad A., Tacconi E., Zimmer J., Liang Y., Gray N.S., Tarsounas M, & Knapp S. (2014). A unique inhibitor binding site in ERK1/2 is associated with slow binding kinetics. Nature chemical biology, 10(10), 853-860.
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6

Inhibition of ERK and BRAF in Cells

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Pharmacological inhibitors, VX-11E (ERK inhibitor) and GSK2118436A (Dabrafenib) were obtained from Chemietek and dissolved in DMSO (Sigma-Aldrich) to a stock concentration of 10 mM. Cisplatin (aqueous) was a kind gift from Matthew Hall, LCB, NCI. Cells were incubated with media at final concentrations of 3μM and 30 μM for 24hrs and 4hrs respectively to determine ERK activity. Inhibitors were used alone and in combination.
Blehm B.H., Jiang N., Kotobuki Y, & Tanner K. (2015). Deconstructing the role of the ECM microenvironment on drug efficacy targeting MAPK signaling in a pre-clinical platform for cutaneous melanoma. Biomaterials, 56, 129-139.
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7

Evaluating EGFR-Targeted Therapies in Cell Lines

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Patient tumor specimens and normal blood were obtained from patients treated at the Massachusetts General Hospital under Institutional Review Board-approved studies. All patients provided written, informed consent, and studies were conducted in accordance with the Declaration of Helsinki.
VACO432 and COLO-320DM cells were obtained from the Massachusetts General Hospital Center for Molecular Therapeutics, which performs routine cell line authentication testing by single-nucleotide polymorphism and short tandem repeat analysis, and were passaged less than six months following receipt. Cells were grown in DMEM/F12 (GIBCO) with 10% FBS and assayed in DMEM/F12 with 5% FBS. Patient-derived cell line (patient 3) was generated and propagated from biopsy material in ACL-4 media (GIBCO) with 10% FBS and assayed in ACL-4 with 5% FBS. Cetuximab and Pantimumab were obtained from the Massachusetts General Hospital Pharmacy and diluted in PBS. Chemical inhibitors from the following sources were dissolved in DMSO: selumetinib, vemurafenib, dabrafenib, trametinib (Selleck Chemicals); Vx-11e (ChemieTek).
Ahronian L.G., Sennott E.M., Van Allen E.M., Wagle N., Kwak E.L., Faris J.E., Godfrey J.T., Nishimura K., Lynch K.D., Mermel C.H., Lockerman E.L., Kalsy A., Gurski JM J.r., Bahl S., Anderka K., Green L.M., Lennon N.J., Huynh T.G., Mino-Kenudson M., Getz G., Dias-Santagata D., Iafrate A.J., Engelman J.A., Garraway L.A, & Corcoran R.B. (2015). Clinical acquired resistance to RAF inhibitor combinations in BRAF-mutant colorectal cancer through MAPK pathway alterations. Cancer discovery, 5(4), 358-367.
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