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SUMO 结合实体（SUBEs）作为肝癌中 SUMO 蛋白酶的浓缩、分离、识别和表征的工具

Journal JoVE
Biochimie

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Journal JoVE Biochimie

SUMO-Binding Entities (SUBEs) as Tools for the Enrichment, Isolation, Identification, and Characterization of the SUMO Proteome in Liver Cancer

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SUMO 结合实体（SUBEs）作为肝癌中 SUMO 蛋白酶的浓缩、分离、识别和表征的工具

DOI:

10.3791/60098-v

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08:29 min

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November 01, 2019

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Fernando Lopitz-Otsoa, Teresa C Delgado, Sofía Lachiondo-Ortega, Mikel Azkargorta, Felix Elortza, Manuel S Rodríguez, María Luz Martínez-Chantar

¹Liver Disease and Liver Metabolism Lab,CIC bioGUNE, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), ²Proteomics Platforms,CIC bioGUNE, ProteoRED-ISCIII, CIBERehd, ³Advanced Technology Institute in Life Sciences (ITAV)-CNRS-IPBS,UbiCARE

Chapitres

00:04Titre
01:58Cell Preparation and Lysis
03:27Tissue Preparation and Lysis
03:54Binding of GST-SUBEs or GST Control to Glutathione-Agarose Beads
04:52GST Pull Down Assay
05:59Identification and Characterization of SUMO Targets
06:36Results: Western Blot and Mass Spectrometry Analysis
07:33Conclusion

Summary

Traduction automatique

在这里，我们提出了一个协议，以丰富、分离、识别和描述通过SUMO结合实体（SUBEs）从小鼠模型获得的从人肝细胞和肝肿瘤中获得的体内由SUMO修饰的蛋白质。

Tags

SUMO-binding Entities SUBEs SUMO Proteome Liver Cancer SUMOylation Protein Isolation Protein Characterization Protease Tandem Repeats SIMs Therapeutic Approach Human Hepatoma Cells Cell Lysis Trypsinization

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