Primary antiviral activity of ALLINIs was observed during virion maturation, where they inhibit IN-RNA interactions [14], we examined the ability of STP0404 to inhibit recombinant IN binding to synthetic TAR RNA using Alpha screen based assay [14]. Briefly, different concentrations of STP0404 were incubated with 100 nM His6 tagged IN in buffer containing 100 mM NaCl, 1 mM MgCl2, 1 mM DTT, 1mg/mL BSA, 25 mM Tris, pH 7.4 at 4°C for 2 hrs. This mixture was then added to the nickel acceptor beads while biotinylated-TAR RNA was added to the streptavidin donor beads. After 2-hrs incubation at 4°C, the RNA mixture was added to the IN-drug mixture and the reading was taken after 1 hr incubation at 4°C by PerkinElmer Life Sciences Enspire multimode plate reader. The IC50 values were calculated by OriginLab software.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.