A ligand-receptor binding assay by receptor immobilization.
Using transferrin-transferrin receptor binding as a model of ligand-receptor binding, we have developed a new and simple binding assay for the solubilized receptor. Solubilized membrane proteins containing transferrin receptor were immobilized by covalent binding to beads having chemical reactive epoxide groups, and then 125I-labeled transferrin was added to the beads. Dose-dependent, ligand-specific, and saturable binding of 125I-labeled transferrin to the immobilized membrane proteins were demonstrated and a Scatchard analysis derived affinity of Kd = 1.8 X 10(-9) M was obtained. These results indicate that the immobilization of receptors onto beads may be useful in a simple binding assay of the solubilized receptor.
Takami
M
M
Division of Biochemical Genetics, Meiji Institute of Health Science, Kanagawa, Japan.
Kasuya
I
I
Tsunoo
H
H
D005260:Female; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis; D008871:Microvilli / Q000032:analysis; D010920:Placenta / Q000032:analysis; D011869:Radioligand Assay / Q000379:methods*; D011990:Receptors, Transferrin / Q000032:analysis; D014168:Transferrin / Q000032:analysis
D008565:Membrane Proteins; D011990:Receptors, Transferrin; D014168:Transferrin