The presence of human seminal plasma proteins in biological stains was demonstrated by an absorption test using anti-human seminal protein rabbit serum. A decreased degree of antibody activity was titrated on a microtiter plate by the agglutination of latex particles coated with human seminal plasma proteins. This method of test was sensitive and highly specific. The latex reagent for this test could be preserved in a refrigerator for over one year without the loss of reactivity.
In 1945-6, Man1' reported the presence of fructose in bull semen. Identification was effected by the preparation of p-methylphenylosazone of fructose. Goto and Ugami2' used later a different technique, paper chromatography, and found to contain only fructose in bull semen. Kubicek and Santavy13' reported that fructose is accompanied with glucose, xylulose, ribose, ribulose and some polysaccharides in the seminal fluids of men and several other animals. Clemont, Glegg and Leblond4' reported that the acrosome of guinea pig spermatozoon contains galactose, mannose, fucose and hexosamines. However, the presence of N-acylaminosugars in free form in human semen has not yet been clarified.Paper chromatography appears to be a sensitive method for the detection of the free N-acylaminosugars in human semen, and it has been applied for that purpose. Preliminary chromatograms on semens from men failed to reveal the presence of the free N-acylaminosugars. However, when the semens were deionized, concentrated and chromatographed, then several N-acylaminosugars appeared. The present paper describes the methods employed for deionization, paper chromatography, and identification of the free N-acylaminosugars in human semen.ExperimenraL Deionization and concentration. From 3 to 5ml of fresh semen from man was dialysed in a cellophan bag against 300-500ml of distilled water at 4-5°C for 24 hours. After dialysis, the dialysate was passed through a column (12mm in internal diameter and 150mm long) of Amberlite IR-120 resin (hydrogen form), 100-200 mesh, and the column was washed with 30-50ml of distilled water. The effluent and washings are combined. The combined solution was then passed through a column (12mm in internal diameter and 150 mm long) of Amberlite IR-4B resin (hydroxyl form), 100-200 mesh, and the column was washed with 30-50m1 of distilled water. The rate of flow of the effluents was 1ml/minute. The effluent and washings are combined. The combined solution was concentrated to about d 2-3ml at 45-50°C under vacuum and the concentrated solution was *1 . Sanban-cho, Chiyoda-ku, Tokyo.
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