Hello,

Try titrating the components, adjusting buffer conditions, progressive mixing, pre-incubation, temperature optimization's, co-factors and additives, and size-exclusion chromatography to prevent precipitation while crystallizing RNA-bound HIV-I Integrase. In order to stabilize the protein-RNA complex, titration entails lowering the concentration of either protein or RNA, watching for precipitation at lower concentrations, and modifying buffer conditions. Pre-incubation provides for a preliminary connection between the components while gradual mixing allows for less firm interactions. By reducing aggregation, temperature optimization's can give the complex more time to develop. Molecular chaperones, certain ions, or tiny molecules are examples of co-factors and additives that can improve the stability of complexes. By removing aggregated or misfolded species, size-exclusion chromatography creates a stable, pure complex that is suited for crystallization investigations.

Regards

Hi Gayatri Munieswaran ,

Thank you for your suggestion.

I tried some of your suggestions such as co-factors, and different buffer components. But I haven't done SEC with once I mix the protein with RNA. I am going to try this and add Arginine and NDSB-201.

Thank you very much!