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Parameters to be calculated
Kd – the dissociation constant The lower the value of Kd the higher the binding affinity Has molar units Values usually in nM range (10-9M) Bmax – number of receptors Often quoted /mg of protein Can be very low fmole/mg (10-15 mole/mg)
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Radioligand binding experiments
Two types Saturation binding Competition Saturation Binding Can calculate BOTH Kd and Bmax Disadvantage – rather expensive – use a lot of radioligand Only calculate Kd Cheaper – use less radioligand
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Saturation binding expt
Preparation of protein Mix with radioligand Incubate Filter Count radioactivity
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