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  • Glioblastoma patient-derived cell-based phenotypic drug screening and identification of possible action mechanisms through proteomic analysis.

Glioblastoma patient-derived cell-based phenotypic drug screening and identification of possible action mechanisms through proteomic analysis.

STAR protocols (2021-10-07)
Young Eun Kim, Hyun Young Kim, Daeyoung Jung, Dukjin Kang, Do-Hyun Nam, Hye Jin Nam, Heeyeong Cho
摘要

Because glioblastoma (GBM) exhibits high heterogeneity, it is desirable to use patient-derived cells from the first stage of screening for GBM drug discovery. Here, we describe a protocol to culture patient-derived GBM cells on the extracellular matrix-coated plates to allow high-throughput screening. Further, we detail approaches to identify the mechanism of action (MOA) of the selected effective drug through proteomics. This protocol will be useful for researchers interested in drug screening and the MOA of drugs. For complete details on the use and execution of this protocol, please refer to Nam et al. (2021).

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Sigma-Aldrich
层粘连蛋白 来源于 Engelbreth-Holm-Swarm 小鼠肉瘤基底膜, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Roche
cOmplete Mini蛋白酶抑制剂Cocktail, Tablets provided in a glass vial
Sigma-Aldrich
碘乙酰胺, BioUltra
Sigma-Aldrich
尿素, powder, BioReagent, Molecular Biology, suitable for cell culture
Sigma-Aldrich
DL-二硫苏糖醇, ≥98% (HPLC), ≥99.0% (titration)
Sigma-Aldrich
氯化钠, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
三乙基碳酸氢铵缓冲液, 1.0 M, pH 8.5±0.1
Sigma-Aldrich
L-半胱氨酸, 97%
Sigma-Aldrich
甲酰胺, ≥99.5% (GC), BioReagent, Molecular Biology