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利用商用仪器和自由开源亮度分析软件对蛋白质的聚齐聚进行无标定的体外定量化
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Biochimica
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JoVE Journal Biochimica
Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software
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利用商用仪器和自由开源亮度分析软件对蛋白质的聚齐聚进行无标定的体外定量化

DOI:
10.3791/58157-v

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08:22 min

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July 17, 2018

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, , , , ,
1Cellular Imaging Group, Wellcome Centre Human Genetics,University of Oxford, 2Division of Structural Biology, Wellcome Centre Human Genetics,University of Oxford, 3Dynamic Structural Virology Group,Biocruces Health Research Centre, 4IKERBASQUE,Basque Foundation for Science

Capitoli

  • 00:04Titolo
  • 00:50FKBP12F36V-mVenus Purification
  • 03:24Preparation of Multiwell Plate Array
  • 04:19Calibration-free Confocal Acquisition
  • 06:26Results: FKBP12 Dimerizes upon Addition of BB Dimerizer Drug
  • 07:23Conclusion

Summary

Traduzione automatica

Traduzione automatica

本协议描述了一种无标定的方法, 用于在荧光涨落光谱的基础上, 利用商用光扫描显微镜定量测定蛋白质的体外齐聚。给出了正确的采集设置和分析方法。

Tags

Keywords: Protein Homo-oligomerizationIn Vitro QuantificationCalibration-freeConfocal MicroscopyBrightness AnalysisProtein-protein InteractionsDrug ScreeningCancer DrugsSmall-molecule InhibitorsFusion InhibitorsProtein DynamicsCell-cell InteractionsFKBP12His6 TagMVenus TagIMACSize-exclusion Chromatography

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