Factors influencing the isolation of membrane immunoglobulins from T and B lymphocytes. 1. Detergent effects and iodination conditions.
The optimal conditions for the extraction and isolation of immunoglobulins from the cell membrane (M-Ig) of T and B lymphocytes radioiodinated with lactoperoxidase were studied. A combination of 10M-urea, 1.5 M acetic acid and the non-ionic detergents Triton X-100 or Nonidet P-40 (NP-40) was found to overcome the tendency for membrane proteins to precipitate after extraction in urea-acetic acid alone. However, detection of T cell Ig was more dependent on detergent concentration than B cell M-Ig. Less T cell Ig was detected when the detergent concentration was increased from 0.1% to 1.0%, whereas the amount of B cell M-Ig was not affected by higher detergent concentrations.
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D001666:Binding Sites, Antibody; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007457:Iodine Radioisotopes; D007784:Lactoperoxidase / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008819:Mice, Nude; D011092:Polyethylene Glycols / Q000494:pharmacology; D011947:Receptors, Antigen, B-Cell / Q000032:analysis / Q000302:isolation & purification*; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
D007457:Iodine Radioisotopes; D011947:Receptors, Antigen, B-Cell; D011092:Polyethylene Glycols; D007784:Lactoperoxidase