Related Experiment Video
Updated: Jul 10, 2025
High Throughput In Vitro Assessment of Latency Reversing Agents on HIV Transcription and Splicing
Published on: January 22, 2019
Protocol for an in vitro assay to study HIV-1 Tat methylation
Daniela Boehm1, Katrin Kaehlcke2, Martina Schnoelzer3
1Gladstone Institute of Virology, University of California San Francisco, San Francisco, CA 94158, USA; Department of Medicine, University of California San Francisco, San Francisco, CA 94143, USA.
Abstract:
A critical virus-encoded regulator of HIV-1 transcription is the Tat protein, which is required to potently activate transcription. Tat is regulated by a wide variety of post-translational modifications. This protocol describes an in vitro assay to study Tat methylation. We describe steps for incorporation of radioactive methyl groups into Tat protein, visualization by gel analysis, Coomassie blue stain, gel drying, and detection by autoradiography. This protocol can also be used to assess methylation in other proteins such as histones. For complete details on the use and execution of this protocol, please refer to Boehm et al. (2023).1.
