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用于神经元细胞贩运研究的管子中纯化和直接单生物素重组BDNF的改进协议
JoVE Journal
Neurociência
Author Produced
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JoVE Journal Neurociência
An Improved Protocol to Purify and Directly Mono-Biotinylate Recombinant BDNF in a Tube for Cellular Trafficking Studies in Neurons
DOI:

13:46 min

July 11, 2020

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Capítulos

  • 00:00Introdcution
  • 00:48Transfection of HEK293 cells
  • 01:49Media Collection and Storage
  • 03:02Media Concentration and Purification
  • 04:58In vitro Mono-biotinylation Reaction
  • 06:01Biotinylation Analysis
  • 06:56Visualization of Retrograde Axonal Transport of BDNF-QD in Live Neurons
  • 07:58Represetative Results
  • 13:17Conclusion

Summary

Tadução automática

含有 Avi 序列 (BDNFAvi) 的重组 BDNF 以经济高效的方式在 HEK293 细胞中生产,并通过亲和色谱进行纯化。然后,BDNFavi 与管中的酶 BirA 直接单生物素化。与市售的BDNF相比,BDNFavi和单生物素化BDNFavi保留了其生物活性。

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