Dear All, Firstly, sorry for the non-crystallography question but I am trying to do a pulldown assay using Cobalt-NTA resin between two DNA replication proteins one of which contains a C-terminal 6x his tag. According to previous genetic studies, these two proteins are involved in replication and many believe there is a complex formed between the two. My question is does anyone know if there is a range of affinities for which a pull down assay is appropriate? And if so is there a reference one could recommend?
Secondly, if there is a weak interaction between two proteins, for example millimolar affinity, does that decrease the chances of co-crystallizing the two interacting? Thank you in advance, Sincerely, James Qiu
