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Abstract Analytical isoelectric focusing in thin layers of polyacrylamide gel was shown by Awdeh, Williamson this has been called the antibody isoelectric spectrum. The usefulness of this method has been extended by employing radioactive hapten to visualize the isoelectric spectrum of anti‐hapten antibodies present in sera. Samples of immune serum are focused, usually using pH 5–8 range Ampholine. Labeled hapten is applied to the surface of the thin layer gel and allowed to combine with focused antibodies; after fixation of the antibody‐hapten complexes, and washing to remove excess hapten, the gel is air dried and a radioautograph taken on X‐ray film. This visualization technique has been shown to be highly specific for antibody against the developing hapten. The sensitivity of detection of antibody is primarily limited only by the specific activity of the radioactive hapten. The visualization of anti‐DNP spectra is described and the extension of the method to other antibodies is discussed. An interpretation of the isoelectric spectrum of antibody present in serum in terms of the number of different antibodies originally synthesized by the animal is presented.
A. R. Williamson (1971) studied this question.