A highly efficient chemical isolation procedure for the rat placental transferrin receptor.
A chemical method for the purification of rat placental transferrin receptor is described. After initial solubilization and concentration by ammonium sulfate precipitation, radioiron-tagged diferric transferrin was added to the dialyzed receptor fraction and subjected to anion-exchange chromatography on DEAE-Sephacel. Elution with a Tris-HCl buffer gradient yields a single fraction of radioactivity containing both free transferrin and the receptor-transferrin as a complex. Further separation of the receptor-transferrin complex from the free transferrin is achieved by gel chromatography on a AcA34-Sepharose 6B separation system. Final purification is obtained by preparative gel electrophoresis in 5% polyacrylamide gels. The receptor was shown to be pure by various methods including HPLC chromatography. The average yield was 20-30 mg receptor-transferrin complex/100 g placental tissue. Because of the purely chemical approach, this method is universally applicable for the isolation of transferrin receptors from various tissues.
Huebers
H A
HA
Department of Medicine, University of Washington, Seattle.
Huebers
E
E
Josephson
B
B
Csiba
E
E
D000645:Ammonium Sulfate; D000818:Animals; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005026:Ethylene Glycols; D005260:Female; D010920:Placenta / Q000032:analysis*; D011247:Pregnancy; D051381:Rats; D011919:Rats, Inbred Strains; D011990:Receptors, Transferrin / Q000302:isolation & purification*
D005026:Ethylene Glycols; D011990:Receptors, Transferrin; C021754:terics; D000645:Ammonium Sulfate