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Agi 5198

Manufactured by Selleck Chemicals
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AGI-5198 is a chemical compound used as a laboratory reagent. It is a specific inhibitor of the IDH1 enzyme. The core function of AGI-5198 is to facilitate the study of the role of the IDH1 enzyme in various biological processes.

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Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions) Wnt agonist 1, by Selleck Chemicals (2 mentions) Mitomycin c, by Selleck Chemicals (2 mentions) Geneprint 10 system, by Promega (2 mentions) Olaparib, by Selleck Chemicals (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Temozolomide, by Selleck Chemicals (1 mentions) Bmn673, by Selleck Chemicals (1 mentions) Resazurin, by Thermo Fisher Scientific (1 mentions) Sodium chloride (nacl), by Thermo Fisher Scientific (1 mentions) Diaphorase from clostridium kluyveri, by Merck Group (1 mentions) Ml309, by Merck Group (1 mentions) Gsk864, by Cayman Chemical (1 mentions) Dl isocitric acid trisodium salt hydrate, by MP Biomedicals (1 mentions) Tris hydrochloride, by Thermo Fisher Scientific (1 mentions) Nicotinamide mononucleotide, by Merck Group (1 mentions) Pdd00017273, by Bio-Techne (1 mentions) Octyl r 2hg, by Merck Group (1 mentions) Doxycycline hyclate, by Merck Group (1 mentions) Temozolomide, by Cayman Chemical (1 mentions)

6 protocols using agi 5198

1

Characterization of Glioma Cell Lines

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All reagents were purchased from Sigma-Aldrich (St Louis, MO) unless otherwise stated. The human astroglial cell line SVG-10B1 was originally established by Dr. Major14 (link),15 (link) and obtained via a material transfer agreement. Human glioblastoma U87 and U373 cells were original purchased from ATCC. All cell lines are free of mycoplasma and authenticated with short tandem repeat (STR) profiling by Johns Hopkins Genetic Resources Core facility using Promega GenePrint 10 system (Madison, WI). SVG cells were cultured in Dulbecco’s Minimum Essential Media (DMEM, Thermo Fisher Scientific, Grand Island, NY) supplemented with 10% fetal calf serum (FCS, Gemini Bio-products, West Sacramento, CA). Cells were incubated in a humidified incubator containing 5% CO2/95% air at 37°C, and passaged every 2–4 days.
The mutant IDH1 inhibitor AGI 5198 was purchased from Selleckchem (Houston, TX) and used to treat the cells at 1.5 μmol/L for 2 weeks; mitomycin C (MMC) at 2.5μg/ml for 4 h, Wnt agonist 1 (Selleckchem) at 1 μmol/L overnight.
Wei S., Wang J., Oyinlade O., Ma D., Wang S., Kratz L., Lal B., Xu Q., Liu S., Shah S.R., Zhang H., Li Y., Quiñones-Hinojosa A., Zhu H., Huang Z.Y., Cheng L., Qian J, & Xia S. (2018). Heterozygous IDH1R132H/WT created by “single base editing” inhibits human astroglial cell growth by downregulating YAP. Oncogene, 37(38), 5160-5174.
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2

Combination Therapy Protocol for Cancer

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SAHA, olaparib, BMN-673, AGI-5198 and temozolomide were all obtained from SelleckChem, and dissolved in DMSO for cell culture experiments. For animal tumor studies, BMN-673 was diluted in a vehicle containing 10% dimethylacetamide, 6% Kolliphor HS 15 and 84% PBS. SAHA was diluted in PBS. Both were administered to mice via oral gavage five days a week, for three weeks. Octyl-2-HG ((2R)-octyl-alpha-hydroxyglutarate) (Cayman Chemical) and Octyl-α-KG (Sigma-Aldrich) were dissolved in DMSO. Horizon Dharmacon ON-TARGETplus Human BRCA1 and RAD51 siRNA were used at 20 nM for 72 hours.
Dow J., Krysztofiak A., Liu Y., Colon-Rios D.A., Rogers F.A, & Glazer P.M. (2021). Vulnerability of IDH1 mutant cancers to histone deacetylase inhibition via orthogonal suppression of DNA repair. Molecular cancer research : MCR.
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3

Isocitrate Dehydrogenase Enzyme Assay

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Tris–hydrochloride, sodium chloride, magnesium chloride hexahydrate, αKG (sodium salt), and resazurin were purchased from Fisher (Hampton, NH). The pH of αKG stocks was adjusted to 7.0 before use. dl-isocitric acid trisodium salt hydrate was purchased from MP Biomedicals (Santa Ana, CA). NADPH (tetrasodium salt) and NADP+ (disodium salt) were purchased from Calbiochem (San Diego, CA). Diaphorase from Clostridium kluyveri and dimethyl sulfoxide (DMSO) were purchased from Sigma–Aldrich (St. Louis, MO). Bovine serum albumin (BSA) was purchased from SeraCare Lifescience (Milford, MA). ML309 was purchased from Sigma, but the manufacturer unable to confirm if the compound is stereospecific or racemic (Sigma–Aldrich, St. Louis, MO). AGI-5198 was purchased from Selleckchem (Houston, TX). GSK864 was purchased from Cayman Chemical Company (Ann Arbor, MI).
Avellaneda Matteo D., Wells G.A., Luna L.A., Grunseth A.J., Zagnitko O., Scott D.A., Hoang A., Luthra A., Swairjo M.A., Schiffer J.M, & Sohl C.D. (2018). Inhibitor potency varies widely among tumor-relevant human isocitrate dehydrogenase 1 mutants. The Biochemical journal, 475(20), 3221-3238.
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4

Characterization of Glioma Cell Lines

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All reagents were purchased from Sigma-Aldrich (St Louis, MO) unless otherwise stated. The human astroglial cell line SVG-10B1 was originally established by Dr. Major14 (link),15 (link) and obtained via a material transfer agreement. Human glioblastoma U87 and U373 cells were original purchased from ATCC. All cell lines are free of mycoplasma and authenticated with short tandem repeat (STR) profiling by Johns Hopkins Genetic Resources Core facility using Promega GenePrint 10 system (Madison, WI). SVG cells were cultured in Dulbecco’s Minimum Essential Media (DMEM, Thermo Fisher Scientific, Grand Island, NY) supplemented with 10% fetal calf serum (FCS, Gemini Bio-products, West Sacramento, CA). Cells were incubated in a humidified incubator containing 5% CO2/95% air at 37°C, and passaged every 2–4 days.
The mutant IDH1 inhibitor AGI 5198 was purchased from Selleckchem (Houston, TX) and used to treat the cells at 1.5 μmol/L for 2 weeks; mitomycin C (MMC) at 2.5μg/ml for 4 h, Wnt agonist 1 (Selleckchem) at 1 μmol/L overnight.
Wei S., Wang J., Oyinlade O., Ma D., Wang S., Kratz L., Lal B., Xu Q., Liu S., Shah S.R., Zhang H., Li Y., Quiñones-Hinojosa A., Zhu H., Huang Z.Y., Cheng L., Qian J, & Xia S. (2018). Heterozygous IDH1R132H/WT created by “single base editing” inhibits human astroglial cell growth by downregulating YAP. Oncogene, 37(38), 5160-5174.
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5

Chemical Compounds for Cell Culture

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The following chemical compounds were used in culture: AGI-5198 (Selleckchem), FK866 (Cayman Chemical), Dimethyl Sulfoxide (Sigma-Aldrich), Doxycycline hyclate (Sigma-Aldrich), Nicotinamide (Sigma-Aldrich), Nicotinamide mononucleotide (Sigma-Aldrich), Octyl-(R)-2HG (Sigma-Aldrich), Olaparib (Selleckchem), PDD00017273 (Tocris), and Temozolomide (Cayman Chemical).
Nagashima H., Lee C.K., Tateishi K., Higuchi F., Subramanian M., Rafferty S., Melamed L., Miller J.J., Wakimoto H, & Cahill D.P. (2020). Poly(ADP-ribose) glycohydrolase inhibition sequesters NAD+ to potentiate the metabolic lethality of alkylating chemotherapy in IDH mutant tumor cells. Cancer discovery, 10(11), 1672-1689.
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6

Cell Line Cultivation and Characterization

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U87, T98G, HT1080 cell lines and U87 IDH1 R132H/þ -mutant isogenic line were obtained from ATCC. U87 and T98G cells were cultured in DMEM (Gibco) supplemented with 10% FBS (Life Technologies), 2 mmol/L L-glutamine, penicillin, and streptomycin (Sigma). HT1080 cells were cultured in Eagle Minimum Essential Medium supplemented with 10% FBS, 2 mmol/L L-glutamine, penicillin, and streptomycin. Normal Human Astrocytes were purchased from Lonza and cultured in Astrocyte Growth Medium BulletKit (Lonza). HCT116 parental and IDH1 knock-in clones (R132H/þ) were obtained from Horizon Discovery and cultured in DMEM supplemented with 10% FBS, 2 mmol/L L-glutamine, penicillin, and streptomycin. All cell lines were tested for Mycoplasma every 3 months and authenticated by short tandem repeat profiling. The number of passages between cells thawing and use in all the experiments was limited in 10 passages. Doxycycline (D9891), 25-Hydroxycholesterol (H1015), and N-acetyl cysteine (A7250) were purchased from MilliporeSigma. AGI-5198 was purchased from Selleck (S7185).
Yang Z., Hu N., Wang W., Hu W., Zhou S., Shi J., Li M., Jing Z., Chen C., Zhang X., Yang R., Fu X, & Wang X. (2022). Loss of FBXW7 Correlates with Increased IDH1 Expression in Glioma and Enhances IDH1-Mutant Cancer Cell Sensitivity to Radiation. Cancer research, 82(3).
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