Radial immunodiffusion quantitation of proteins by an antibody overlay technique.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to R J Luby.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Up until now it has been assumed that the protease-binding property of alpha1-protease inhibitor (alpha1PI) was destroyed by acid starch gel electrophoresis (pH 4.9). Analyses on acid starch gel blocks for pH and conductivity changes during and following a typical electrophoretic run showed that it was unlikely that the separating alpha1PI would be exposed to pH values lower than 6.2, and that the allele products, following the passage of the buffer front, were in an environment of constant pH(6.3), extremely low conductivity and high field strength. These results strongly suggested the likelihood that alpha1-PI would be chemically and physically unchanged as a result of exposure to acid starch gel electrophoresis. In order to test this likelihood, human serum was electrophoretically separated in acid starch gel and following electrophoresis, was immersed in 0.1 M diethylbarbiturate buffer, pH 8.6, containing 20 mug/ml of pancreatic elastase. The pH-adjusted (8.15) and elastase-impregnated starch gel layer was superimposed on hemoglobin-agar for 2.5 h at 37 degrees C followed by immersion of the hemoglobin-agar layer in 1% NaCl overnight, distilled water for 2 h, drying under filter paper and staining. The results showed zones of undigested hemoglobin indicating, unequivocally, that the separated alpha1PI allele products are capable of forming complexes with proteases and that alpha1PI is not inactivated following exposure to acid starch gel electrophoresis. Densitometric analysis of the transparent stained zones on a clear agar gel background offers an alternative to analysis of the acid starch gel-separated zones by antigen-antibody crossed electrophoresis and as such is suitable for identification of alpha1-protease inhibitor phenotypes. Further, the method is specific for alpha1PI and a densitometric scan provides direct information relative to the protease-binding capacity of the sample as well as the contribution of each alpha1PI allele product to that capacity.
Following ovulatory control cycles in four celibate, healthy women, 27 to 34 years of age, basal body temperatures (BBT) were obtained in three cycles, utilizing a transistorized transvaginal telemeter. The electronic signals were collected on an automatically activated cassette receiver and later translated into temperature readings. The results were compared with values obtained simultaneously with a standard metabolic oral thermometer. The vaginal temperatures were consistently lower than the oral readings and offered a more sensitive and reproducible method of obtaining BBT. However, this system is very complex, requiring well-informed, highly motivated patients, and should be viewed currently as an investigative tool.