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通过 SHP2 (PTPN11) 磷酸酶抑制剂评估细胞目标参与度
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JoVE Journal
Cancer Research
Assessing Cellular Target Engagement by SHP2 (PTPN11) Phosphatase Inhibitors
Please note that all translations are automatically generated.
Click here for the English version.
通过 SHP2 (PTPN11) 磷酸酶抑制剂评估细胞目标参与度
DOI:
10.3791/61457-v
•
08:45 min
•
July 17, 2020
•
Lester J. Lambert
,
Celeste Romero
,
Douglas J. Sheffler
,
Maria Celeridad
,
Nicholas D. P. Cosford
,
Lutz Tautz
1
Cancer Metabolism & Signaling Networks Program, NCI-Designated Cancer Center
,
Sanford Burnham Prebys Medical Discovery Institute
Chapters
00:04
Introduction
01:04
Transfection of HEK293T Cells
02:36
Incubation of Cells with SHP2 Inhibitors
05:11
Isothermal or Thermal Profile Gradient Heat Pulse
06:01
Results: Cellular Target Engagement of Allosteric SHP2 Inhibitors with SHP2-WT and SHP2-E76K Proteins
08:03
Conclusion
Summary
Automatic Translation
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Automatic Translation
评估候选抑制剂在完整细胞中参与目标的能力对药物的发现至关重要。该协议描述了一个384井格式的细胞热移检测,可靠地检测针对野生型SHP2或其致癌变异的抑制剂的细胞靶接触。
Tags
SHP2
Phosphatase
Cell Signaling
Cancer Therapy
Small Molecule Inhibitors
Target Engagement
Cell-based Assay
HEK293 T-cells
Transfection
384-well Plates
Liquid Handling
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