FAQ: How much lysate can be loaded onto a single NEBExpress Ni Spin Column?

Each column can hold up to 500 µl of lysate per application and provides approximately 1 mg of available binding capacity. The binding efficiency can vary significantly between targets and is dependent on the size of the protein and the accessibility of the His-tag.  It is important to estimate the expression level of your target protein, this can be done by running both the lysate and a standard (of known concentration) for quantitation using SDS-PAGE.