Differential radiolabelling of lymphocyte membrane alloantigens and immunoglobulins: variation of H2O2 concentration during lactoperoxidase catalyzed cell surface radio-iodination.

The effect of H2O2 concentration on lactoperoxidase catalyzed cell surface radio-iodination and subsequent isolation of Murine splenic lymphocyte Ia, H-2K and Lyb-3 surface antigens and membrane immunoglobulins was studied. For most membrane polypeptides analyzed 0.3 mM H2O2 proved to be optimal for the recovery of radiolabelled antigens from detergent lysates of labelled cells by immunoprecipitation. Marked variations among surface antigens and membrane immunoglobulin polypeptide chains were observed for the iodination and recovery of these proteins above and below the optimal peroxide concentration. The results suggest that cell surface radio-iodination conditions should be standardized to the requirements of the particular membrane protein being studied. The differential iodination and recovery of discrete membrane components above and below optimal conditions may prove useful in the analysis of surface membrane protein structure and membrane association.