Radioimmunoassay of prostaglandin F2 alpha using sephadex G-75 gel chromatography.
The development of a "bound-free" separation technique and its application to the radioimmunoassay of prostaglandin F2 alpha is described. The method is simple, rapid, free of non-specific binding and could be performed either at 4 degrees C C or at room temperature. A total of 100 tubes could be subjected to "bound-free" separation in 30 min at 4 degrees C. The bound fraction is collected directly into scintillation vials. The total column length was 9.5 cm, of which the bed volume was 2.5 ml. The PGF2 alpha radioimmunoassay incubation volume of 0.3 ml when bedded in required 1.4 ml of elution buffer to elute the antibody-bound fraction. The free fraction was washed out with 4.0 ml of buffer and the columns were ready for further use. A standard curve of high sensitivity (5 pg) and good reproducibility (CV%: intra-assay = 6.54; inter-assay = 9.68) was obtained.
D000936:Antigen-Antibody Complex / Q000032:analysis; D002850:Chromatography, Gel / Q000379:methods; D006801:Humans; D011460:Prostaglandins F / Q000032:analysis* / Q000276:immunology; D011863:Radioimmunoassay / Q000379:methods
D000936:Antigen-Antibody Complex; D011460:Prostaglandins F