A simple method for the determination of affinity and binding site concentration in receptor binding studies

Biochim Biophys Acta. 1977 Nov 1;470(3):412-23. doi: 10.1016/0005-2736(77)90132-8.


In ligand binding studies, it is often difficult to apply kinetic analyses because of an uncertainty in experimental data obtained at high ligand concentrations. Under such circumstances, Kd value (an index of the affinity) and the binding site concentration may be estimated more accurately from the binding of a fixed concentration of labelled ligand observed in the presence of various concentrations of the non-labelled ligand, if the fraction of both labelled and non-labelled ligand bound is small. When there is no cooperative effect of the ligand binding, the Kd value may be calculated by subtracting the concentration of the labelled drug from the concentration of the non-labelled drug to cause a 50% reduction of the saturable binding of the labelled drug. From above values, the binding site concentration may be calculated. The proposed method is capable of examining the cooperativity of the ligand binding, the labelled drug concentration and the specific radioactivity of the labelled drug and does not require large amounts of the labelled drug.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Animals
  • Binding, Competitive
  • Brain / metabolism*
  • Kinetics
  • Male
  • Mathematics
  • Ouabain / metabolism*
  • Potassium / metabolism
  • Rats
  • Receptors, Drug / metabolism*
  • Sodium / metabolism


  • Receptors, Drug
  • Ouabain
  • Sodium
  • Adenosine Triphosphatases
  • Potassium