Abstract:t-Zeatin (t-Z) and isopentenyladenosine (iPA) occur naturally as highly active plant cell division regulators, t-Z-Sepherose-4B and iPA-Sepherose-4B affinity column were constructed to isolate and purify the cytokinin-binding proteins from etiolated hypocotyl of Phaseolus vulgaris. Two kinds of cytokinin-binding proteins were obtained. One was 15.5 kD in molecular weight (named ZBP) with only one peptide. The other (named IBP), 165 kD in molecular weight, contained two different subunits (40 kD and 43 kD respectively). The binding activity of ZBP was tested and the dissociation constant (Kd) was determined to be 3.2 × 10-7 mol/L. There was one binding site for t-Z in each molecule of ZBP.