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基于刀豆球蛋白A的沉降测定法，用于测量葡聚糖磷酸酶的底物结合
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Bioquímica
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JoVE Journal Bioquímica
Concanavalin A-Based Sedimentation Assay to Measure Substrate Binding of Glucan Phosphatases
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基于刀豆球蛋白A的沉降测定法，用于测量葡聚糖磷酸酶的底物结合

DOI:
10.3791/64700-v

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09:07 min

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December 23, 2022

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, , , ,
1Department of Chemistry,Skidmore College

Capítulos

  • 00:04Introduction
  • 01:03Preparation of ConA‐Sepharose Beads and ConA‐Sepharose: Amylopectin Beads
  • 02:19Starch Excess4 (SEX4) Protein Binding with ConA‐Sepharose: Amylopectin Beads
  • 03:41Running SDS‐PAGE Gels
  • 04:27Western Blotting for Chemiluminescence Detection and Data Analysis
  • 06:28Results: Binding Affinity of SEX4 to ConA‐Sepharose‐Amylopectin Beads
  • 08:18Conclusion

Summary

Tadução automática

Tadução automática

该方法描述了一种基于凝集素的 体外 沉降测定，以量化葡聚糖磷酸酶和支链淀粉的结合亲和力。该共沉降测定法对于测量葡聚糖磷酸酶底物结合是可靠的，可应用于各种可溶的葡聚糖底物。

Tags

Concanavalin ASedimentation AssayGlucan PhosphatasesSubstrate BindingCarbohydrate-protein InteractionsSEX4 MutantsAmylopectinBinding BufferCentrifugationProtease Inhibitor

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