Batch or mixing method? - (Jun/19/2008 )

Which way is more preferred for binding to Ni-NTA column? The method where you add the lysate to the Ni resin and let it mix for a couple hours or the one where you just add it to the column and let it drain through slowly? Im only getting like 3 mg of protien from my workup but some if it goes through into the flow through so im just wondering if some didnt bind. Thanks.

Hi

Its depend on what ur looking for. Letting the resin mix with the protein is preferable for binding

but it also bind undesired protein (contaminating proteins) and only do it for like 40 to 60 min at max.

if ur getting "nice" quantities of the protein with contaminations, it is better to use the batch method

(personally i dont like it)

bye