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Efficient and Site-specific Antibody Labeling by Strain-promoted Azide-alkyne Cycloaddition
JoVE Journal
Biyokimya
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JoVE Journal Biyokimya
Efficient and Site-specific Antibody Labeling by Strain-promoted Azide-alkyne Cycloaddition

Efficient and Site-specific Antibody Labeling by Strain-promoted Azide-alkyne Cycloaddition

DOI:
10.3791/54922-v

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09:06 min

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December 23, 2016

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, , ,
1Kimya Bölümü,Sogang University

Bölümler

  • 00:05Başlık
  • 00:42Culture Preparation
  • 02:10Expression and Purification of HerFab-L177AF
  • 04:30Conjugation of Purified HerFab-L177AF with Alkyne Probes Using Strain-promoted Azide-alkyne Cycloaddition (SPAAC) and Purification of Labeled HerFab
  • 05:52SDS-PAGE Analysis of Labeled HerFab
  • 07:03Results: Site-specific Antibody Labeling by SPAAC
  • 08:07Conclusion

Özet

Otomatik Çeviri

Otomatik Çeviri

Here, we present a protocol to site-specifically introduce chemical probes into an antibody fragment by genetically incorporating an azide-containing amino acid, and subsequently coupling the azide with a chemical probe by strain-promoted azide-alkyne cycloaddition (SPAAC).

Etiketler

Antibody LabelingSite-specificAzide-alkyne CycloadditionProtein ModificationMühendislikConjugationE. ColiPlasmidElectroporationArabinoseCell LysisParaperiplasmic Extraction

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