A simple method for the radioactive iodination of CD4 molecules.
A method for the vectorial radioiodination of CD4 and other membrane proteins having few or no tyrosine residues in the extracytoplasmic domains is described. Incubation of the cells with sulfosuccinimidyl (hydroxyphenyl) propionate (sulfo-SHPP), a water-soluble derivative of the Bolton-Hunter reagent results in the coupling of hydroxyphenyl groups to free amino groups of cell surface proteins and these groups are then vectorially radioiodinated using 1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril (Iodogen). The method is highly efficient, gentle, fast, simple, and does not require the previous radiolabelling of the Bolton-Hunter reagent.
Portolés
P
P
Department of Pathology, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06510.
Rojo
J M
JM
Janeway
C A
CA
D000818:Animals; D001345:Autoradiography; D015704:CD4 Antigens / Q000378:metabolism*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D007457:Iodine Radioisotopes / Q000378:metabolism*; D008722:Methods; D011233:Precipitin Tests; D013388:Succinimides; D014508:Urea / Q000031:analogs & derivatives
D015704:CD4 Antigens; D007457:Iodine Radioisotopes; D013388:Succinimides; C017730:N-succinimidyl-3-(4-hydroxyphenyl)propionate; C012385:1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril; D014508:Urea