Measurement of IgE antibodies by the radioallergosorbent test. I. Technical considerations in the performance of the test.
Since its introduction by Wide, Bennich, and Johansson in 1967, the radioallergosorbent test (RAST) has been increasingly used for measurement of specific IgE antibodies. In this study we have varied the conditions during the performance of the RAST and have investigated the effect of altering: (1) the speed of rotation of tubes, (2) the number of washes in both steps of the test, (3) the length of rotation of tubes, (4) the temperature, (5) the conditions during centrifugation, (6) the quantity of antibody to IgE in the second step of the test, (7) the composition of buffers in the reactions, (8) the type of tube employed in the test, (9) the quantity of antigen employed during the coupling procedure, and, finally, (10) the iodine isotope, 125I or 131I, used to radiolabel anti-IgE. Of these variables the most significant were the quantity of anti-IgE used in the second step of the test and the quantity of antigen used in the coupling procedure. Other variables of importance were the choice of radioiodine, whether the tubes were rotated or not, the type of tube used, and finally the diluent employed.
D000485:Allergens / Q000302:isolation & purification; D000818:Animals; D000906:Antibodies / Q000032:analysis*; D002482:Cellulose; D006382:Hemadsorption; D007073:Immunoglobulin E / Q000032:analysis*; D007457:Iodine Radioisotopes; D008722:Methods; D011058:Pollen; D012756:Sheep / Q000276:immunology; D013696:Temperature
D000485:Allergens; D000906:Antibodies; D007457:Iodine Radioisotopes; D007073:Immunoglobulin E; D002482:Cellulose