![]() All rights reserved.ĬAPILLARY ISOELECTRIC FOCUSING ( CIEF) FOR THE CHARACTERIZATION OF HUMIC SUBSTANCES The results suggest that the i CIEF technique can be used to accurately resolve and quantitate amylase isozymes in a rapid and automated fashion, and that gel-filtration should be applied to saliva samples beforehand to allow for optimal purification and characterization. Furthermore, saliva secreted from the parotid gland proved to have a higher amylase content compared to either secretions from the submandibular/sublingual complex, or whole saliva, as well as amylase enhancement under stimulation. In addition, a high level of precision was apparent, and the relative standard deviation for replicates was less than 0.5% for pIs (peak positions) and below 10% for peak area. Good agreement existed between the observed isoelectric points and the values reported in the literature. Salivary α-amylase separated into two major peaks before sample treatment while both filtration methods and starch affinity interaction of salivary amylase enhanced the resolution of isozymes, desalting with gel-filtration displayed the best recovery and the highest resolution of isozymes. Notably, three sample preparation methods were examined: ultrafiltration, gel-filtration, and starch affinity interaction with salivary amylase. Capillary isoelectric focusing coupled with UV whole column imaging detection (i CIEF) was used to develop a robust protocol to characterize salivary α-amylase collected from various glands. ![]() While gel electrophoresis is generally used for salivary analysis, we employed the capillary isoelectric focusing technique to allow for a rapid, automated mode of electrophoresis. ![]() ![]() Saliva is an easily collected biological fluid with potentially important diagnostic value. Zarabadi, Atefeh S Huang, Tiemin Mielke, John G ![]() Capillary isoelectric focusing with whole column imaging detection (i CIEF): A new approach to the characterization and quantification of salivary α-amylase. ![]()
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