Lactoperoxidase catalyzed radioiodination of cell surface immunoglobulin: incorporated radioactivity may not reflect relative cell surface Ig density.
Rabbit and mouse splenic lymphocytes were radioiodinated by the lactoperoxidase technique, extracted with non-ionic detergent, immunoprecipitated with high titered rabbit anti-kappa antisera, and compared by SDS-PAGE. Mouse sIg peaks were reproducibly larger in size than rabbit sIg peaks (often greater than 10 times). Neither differences in incorporation of label into the rabbit cell surface, nor differences in average sIg density explain this result. Total TCA-precipitable radioactivity was similar in each species. Estimation of the relative amounts of sIg in the mouse and rabbit showed similar average sIg densities. Differences in detergent solubility, proteolytic lability, or antisera used also do not adequately account for this difference. Thus, these data indicate that radioactivity incorporated after lactoperoxidase catalyzed cell surface radioiodination may not reflect cell surface Ig density. Conclusions about cell surface density based upon relative incorporation of radioactivity should be confirmed by other approaches.
D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007457:Iodine Radioisotopes; D007784:Lactoperoxidase / Q000494:pharmacology*; D051379:Mice; D010544:Peroxidases / Q000494:pharmacology*; D011817:Rabbits; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D013205:Staphylococcal Protein A / Q000276:immunology; D013425:Sulfanilic Acids / Q000378:metabolism
D007457:Iodine Radioisotopes; D011947:Receptors, Antigen, B-Cell; D013205:Staphylococcal Protein A; D013425:Sulfanilic Acids; D007784:Lactoperoxidase; D010544:Peroxidases