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JoVE Journal

Genetics

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Analysis of Cap-binding Proteins in Human Cells Exposed to Physiological Oxygen Conditions

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10:40 min

December 28th, 2016

December 28th, 2016

7,678 Views

0:05

Title

1:03

Subculturing Cells and Physioxic Exposure

1:57

Cell Lysis

5:06

Cap-binding Assay

8:44

Results: Analysis of Cap-binding Ability in Response to Oxygen of eIF4E and eIF4E2 in an m7GTP Affinity Column

9:50

Conclusion

Transcript

The overall goal of this experiment is to capture cap-binding proteins in conditions of low oxygen. This method utilizes an agarose linked seven methyl-guanosine cap analog to investigate cap-binding factors and their interacting partners. This me

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Here, we present human cell culture protocols to analyze translation initiation factors that bind the 5' cap of mRNA during physiological oxygen conditions. This method utilizes an Agarose-linked m7GTP cap analog and is suitable to investigate cap-binding factors and their interacting partners.

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