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滴定 ELISA 法测定受体配体相互作用的离解常数
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Titration ELISA as a Method to Determine the Dissociation Constant of Receptor Ligand Interaction
DOI:

12:38 min

February 15, 2018

Chapters

  • 00:04Title
  • 00:45Immobilization of the Receptor (Integrin α2A-domain) to a Microtiter Plate
  • 02:09Preparation of a Serial Dilution Row of the Ligand (Rhodocetin)
  • 03:25Binding of Ligand at Different Concentrations to Immobilized Receptor
  • 05:03Quantification of Receptor-bound Ligand by ELISA
  • 07:06Evaluation of the Titration Signals
  • 09:30Results: Rhodocetin Binds with Lower Affinity to Mutant Integrin α2A-domain
  • 10:58Conclusion

Summary

Automatic Translation

描述了执行滴定 ELISA 的详细协议。此外, 提出了一种新的评价滴定 ELISAs 的方法, 并获得了可溶性配体对微量滴定板固定化受体的离解常数。

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