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从脂质绑定蛋白和过敏原中去除和替换内源性利根

JoVE Journal
Bioquímica

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JoVE Journal Bioquímica

Removal and Replacement of Endogenous Ligands from Lipid-Bound Proteins and Allergens

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从脂质绑定蛋白和过敏原中去除和替换内源性利根

DOI:

10.3791/61780-v

•

09:09 min

•

February 24, 2021

•

Alexander C. Y. Foo, Peter M. Thompson, Geoffrey A. Mueller

¹Nuclear Magnetic Resonance Group,National Institute of Environmental Health Sciences

Capítulos

00:04Introduciton
01:13Endogenous Lipid Removal
02:42Reconstitution of Apo- and Cargo-loaded Bla g 1
05:07Phospholipid Cargo Removal/Loading Confirmation
06:22Results: Representative Bla g 1 Purification, Lipid Removal Verification, and Recovery
08:18Conclusion

Summary

Tadução automática

该协议描述了从过敏原中去除内源性脂质，并通过逆相HPLC与热退化用用户指定的配体替换它们。 ³¹P-NMR 和圆形二分法允许快速确认配体去除/装载，并恢复原生过敏原结构。

Tags

Keywords: Endogenous Ligands Lipid-bound Proteins Allergens Reverse-phase HPLC Thermal Annealing Solubilization Bla G 1 De-lipidation Lyophilization Protein Yield

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