In article <2q8u89$skp at news.iastate.edu>, yqhuang at iastate.edu () writes:
Have tried deionized water or 10-20% ethylene glycol for elution? It is also
sometimes helpful to allow the column to "steep" in the elution buffer for
awhile. Other than that, you might need to use a lower affinity Ab. Regarding
purity, without knowing more details about your procedure its hard to say.
Some matrices have problems with nonspecific binding that can be resolved by
washing the column with relatively mild elution conditions (ex: high then low
salt) prior to elution of the protein of interest. Another possibility is your
protein G, which is known to bind things other than IgG. You might try
directly coupling your antibody to a preactivated matrix (NHS usually works
well); I've had good luck with sepharose-NHS. Pharmacia has a variety of
chemistries available.
