Antibody isoelectric spectra. Analysis of the heterogeneity of antibody molecules in serum by isoelectric focusing in gel and specific detection with hapten

摘要: Analytical isoelectric focusing in thin layers of polyacrylamide gel was shown by Awdeh, Williamson & Askonas (Nature 1968. 219: 66) to be capable resolving a specifically purified antibody into pattern discrete bands; this has been called the spectrum. The usefulness method extended employing radioactive hapten visualize spectrum anti-hapten antibodies present sera. Samples immune serum are focused, usually using pH 5–8 range Ampholine. Labeled is applied surface layer and allowed combine with focused antibodies; after fixation antibody-hapten complexes, washing remove excess hapten, air dried radioautograph taken on X-ray film. This visualization technique highly specific for against developing hapten. sensitivity detection primarily limited only activity anti-DNP spectra described extension other discussed. An interpretation terms number different originally synthesized animal presented.