Identification of Small Molecule-binding Proteins in a Native Cellular Environment by Live-cell Photoaffinity Labeling

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12.5K Views

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10:49 min

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September 20th, 2016

DOI :

10.3791/54529-v

September 20th, 2016

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Sarah A. Head¹, Jun O. Liu¹^,²

¹Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, ²Department of Oncology, Johns Hopkins University School of Medicine

Podsumowanie

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Keywords Small Molecule

Binding Proteins

Photoaffinity Labeling

Live cell

Molecular Mechanism

Drug Target

Protein Identification

Native Cellular Environment

Rozdziały w tym wideo

0:05

Title

0:43

Preparation of Cultured Cells

1:23

Treatment of Cells and Photocrosslinking

4:34

Attachment of Fluorescent Tag by Click Chemistry for Visualization of Labeled Proteins

5:48

Attachment of Biotin Tag by Click Chemistry for Affinity Purification of Labeled Proteins

8:54

Results: Live-cell Photoaffinity Labeling Identifies a Major Itraconazole-binding Protein

10:14

Conclusion

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