The BT474 cell line was maintained in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. The SKBR3 cell line was maintained RPMI1640 supplemented with 10% FBS and 1% penicillin/streptomycin. All cell lines were cultured in humidified incubation at 37 °C with 5% CO2. Lapatinib was purchased from Santa Cruz (Dallas, TX, USA), and 17DMAG-HCL was purchased from Selleck Chemicals (Houston, TX, USA). Stock solutions were prepared with dimethyl sulfoxide (DMSO) and stored at −20 °C. Lapatinib and 17DMAG for animal experiments were purchased from LC laboratories. Lapatinib was dissolved in water with 0.5% carboxymethylcellulose, 1.8% sodium chloride, and 0.4% Tween 80. 17 DAMG was dissolved in water with 0.8% NaCl.
17 dmag
Manufactured by LC Laboratories
Sourced in United States
17-DMAG is a small molecule inhibitor of Heat Shock Protein 90 (HSP90). It is used in research applications to study the role of HSP90 in cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
17 aag, by LC Laboratories (2 mentions)
Lapatinib, by Santa Cruz Biotechnology (1 mentions)
Lapatinib, by LC Laboratories (1 mentions)
Rifabutin, by Merck Group (1 mentions)
17 aag, by Merck Group (1 mentions)
Nvp auy922, by Selleck Chemicals (1 mentions)
Biib021, by Selleck Chemicals (1 mentions)
Gedunin, by Bio-Techne (1 mentions)
Celastrol, by Bio-Techne (1 mentions)
Radicicol, by AG Scientific (1 mentions)
Alamar blue, by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Bleomycin, by Merck Group (1 mentions)
C57bl 6 mice, by Charles River Laboratories (1 mentions)
Flexivent system, by SCIREQ (1 mentions)
6 protocols using 17 dmag
1
Culturing HER2+ Breast Cancer Cell Lines
2
Targeting HSP90 in Immunized and Transplanted Mice
3
Melanoma Cell Culture and Inhibitors
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Culture conditions and the cell lines have been previously described [12] (link). The B16-BL6 murine melanoma cell line was provided by Dr. Andrew Hurwitz [24] (link). Human cell lines were obtained from the ATCC, (Manassas, VA) (including MALME-M3, MM96L, A375, 453A, MM455, Roth, H59-44T, Mel-Juso, JURKAT), or from a patient at the Massashusetts General Hospital (MU89 and MU-X (derived from MU89 as previously described [6] (link), [11] (link)). Construction and culture of the J.RT3-T3.5 cell line expressing a Melan-A/MART-1 specific TCR has been described [15] (link). IFN-β-1a (Avonex) was obtained from Biogen-Idec (Cambridge, MA). The iHsp90s were obtained from the following sources: Radicicol (A.G. Scientific, San Diego, CA); Novobiocin (BioMol, Plymouth Meeting, PA); 17-DMAG (LC laboratories, Woburn, MA); 17-AEP (InVivoGen, San Diego, CA); Rifabutin, PU-H71, and 17-AAG (Sigma, St. Louis, MO); Gedunin, CCT018159, and celastrol (Tocris, Ellisville, MO); and NVP-AUY922 and BIIB021 (Selleck Chemicals, Houston, TX).
4
Cytotoxicity Screening of Antileishmanial Compounds
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Axenic L. amazonensis promastigotes in logarithmic phase were distributed on 96-well plates (4 × 105 parasites/well) in Schneider’s complete medium and treated with GA (Invivogen, San Diego, CA, USA), 17-AAG (LC Laboratories, Woburn, MA, USA) or 17-DMAG (LC Laboratories, Woburn, MA, USA) in 12-step serial dilutions (1:2) at an initial concentration of 2 μM. After 72 h of treatment at 24 °C, parasites were incubated with 10% Alamar Blue® (Invitrogen, Carlsbad, CA, USA) for 1 h30 min. Optical density (OD) was then read at the wavelengths of 570 and 600 nm in a spectrophotometer (SPECTRA Max 340 PC) to determine cell viability, expressed in percentage terms, used to calculate the IC50 values. These were determined by applying sigmoidal regression to the concentration-response curves using GraphPad Prism v5.030 (link) from at least three independent experiments performed in triplicates.
5
Cell Culture of Breast Cancer Cell Lines
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MDA-MB-231 was purchased from The Bioresource Collection and Research Center in Taiwan (Hsinchu, Taiwan) and was maintained in DMEM medium containing 10% fetal bovine serum (FBS). AS-B244 was obtained from Alice L. Yu in Institute of Stem Cell and Translational Cancer Research, Chang Gung Memorial Hospital at Linkou and Chang Gung University (Taoyuan, Taiwan) and was maintained in MEMα medium containing 10% FBS and 5 µg/mL insulin (Sigma-Aldrich, St. Louis, MO, USA). 17-DMAG was purchased from LC Laboratories (Woburn, MA, USA) and dissolved in dimethyl sulfoxide (DMSO) as a stock of 50 mM.
6
Bleomycin-Induced Lung Fibrosis Model
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Adult male 12-week-old C57BL/6 mice (20-25 g body weight) were obtained from Charles River Laboratories (Sulzfeld, Germany). Both the university animal care committee and the Federal Authorities for Animal Research of the Regierungspraesidium Giessen (Hessen, Germany) approved the study protocol (GI20/10, 59/2012). Mice were given bleomycin (3.5 units•kg -1 in saline; Sigma-Aldrich, Munich, Germany) or saline alone via orotracheal instillation. From day 7 to day 21, mice received 17-DMAG (17dimethylaminoethylamino-17-demethoxygeldanamycin; LC Laboratories, Woburn, MA, USA) 10 mg•kg -1 or 25 mg•kg -1 in saline or saline alone via oral gavage every 2 days.
On day 21, mice were scanned using micro-computed tomography (SkyScan, Kontich, Belgium) followed immediately by fluorescence molecular tomography (FMT; VisEn Medical, Bedford, MA, USA) to assess matrix metalloproteinase (MMP) activity. In addition, physiological measurements were performed on day 21 using the FlexiVent system (SCIREQ, Montreal, Canada), bronchoalveolar lavage fluid (BALF) was collected and lungs were fixed for histology or snap-frozen after euthanisation.
On day 21, mice were scanned using micro-computed tomography (SkyScan, Kontich, Belgium) followed immediately by fluorescence molecular tomography (FMT; VisEn Medical, Bedford, MA, USA) to assess matrix metalloproteinase (MMP) activity. In addition, physiological measurements were performed on day 21 using the FlexiVent system (SCIREQ, Montreal, Canada), bronchoalveolar lavage fluid (BALF) was collected and lungs were fixed for histology or snap-frozen after euthanisation.
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