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正确的蛋白浓度的动力学结构与功能分析和亲和力测定的重要性
JoVE Journal
Biologia
This content is Free Access.
JoVE Journal Biologia
The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis
DOI:

19:16 min

March 17, 2010

Capítulos

  • 00:00Título
  • 01:38Introduction
  • 02:30Experimental Design
  • 03:58Immobilization of the Ligand Papain for CFCA Analysis
  • 07:34Determining Cystatin B Concentration Using the CFCA Assay
  • 11:07Measuring the Kinetics of Cystatin B binding to Papain
  • 17:13CFCA assay and Kinetics of Cystatin B binding to Papain Representative Results
  • 18:47Conclusion

Summary

Tadução automática

我们采用无标记蛋白质相互作用分析利用Biacore X100几个胱抑素乙突变体的结合,木瓜蛋白酶,通过动力学特性的结构与功能分析。无标定浓度分析中心(CFCA)的措施,而不需要为标准曲线与保留结合活性的蛋白浓度。我们表明,浓度使用CFCA增加的动力学分析的可靠性,并能够可靠地确定,即​​使重组蛋白的活性降低,动力学常数确认。

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