A one-step procedure for purification of bovine mammary epithelial cell CD36.
A simple one-step protocol for the purification of the integral membrane protein CD36 from milk-fat-globule membranes of bovine mammary epithelial cells is described. Nonionic detergent extracts of membrane were chromatographed on hydroxylapatite and pure CD36 was eluted with 1 M NaCl. Other proteins of the milk-fat-globule membrane were eluted after CD36 with phosphate buffer. Human platelet CD36 bound to hydroxylapatite only after neuraminidase treatment. CD36 is an extremely hydrophobic and highly glycosylated protein and previous purification procedures have required multiple steps. Chromatography of CD36 on hydroxylapatite provides a simple and quick method of purification which does not sacrifice yield.
Greenwalt
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Holland Laboratory, American Red Cross, Rockville, Maryland 20855.
D000818:Animals; D015703:Antigens, CD / Q000302:isolation & purification*; D018955:CD36 Antigens; D002417:Cattle; D002853:Chromatography, Liquid; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D004848:Epithelium / Q000737:chemistry; D006801:Humans; D006882:Hydroxyapatites; D008321:Mammary Glands, Animal / Q000737:chemistry*; D009439:Neuraminidase / Q000737:chemistry; D010980:Platelet Membrane Glycoproteins / Q000302:isolation & purification*
D015703:Antigens, CD; D018955:CD36 Antigens; D006882:Hydroxyapatites; D010980:Platelet Membrane Glycoproteins; D017886:Durapatite; D009439:Neuraminidase