The only way to know for sure is to try it. However, if you have a working method starting at 2 M, you might as well stay with it, unless there is a good reason to lower the starting ammonium sulfate concentration, such as cost or protein stability.

Surely you can load the sample at 1.6 or 2.0 M ammonium sulfate conc and perform elution at 1.4 M or lower. Go ahead.

Hi sir,

If you can elute out the protein solution with 1.4 M Ammonium sulfate salt, then surely try to evaluate the process run with lesser concentration of ammonium sulfate also. Ideally it should not create any binding issues in lower concentration.

So just try to evaluate it.

THe suggestions given by Prof. Adam, Prof Shamsher, and Dr. Mihir are spot on; You may have to try it out at varying concentrations for higher yield of the protein. Alternatively, there are online softwares in which, you could just copy paste your protein's sequence. Based on this, your optimum buffer ranges could be arrived at (column specific). The software would give you a rough idea; Practically performing them would give you the precise concentration.

thank you Adam B Shapiro i will definitely give it a try.

Thank you Shamsher S. Kanwar and Mihir Ranjan Sahoo for your valuble suggestion

Navnit Kumar Ramamoorthy can you please elaborate which software can predict buffers for Hydrophobic chromatography or buffer optimum for protein.