Yao Zhao1, Kaiyong Chen2, Haining Li3, Huiyi Wu4. 1. Medical Technology Institute of Xuzhou Medical University, Xuzhou, 221004, China. 2. Department of Clinical Laboratory, People's Hospital of Guanyun, Lianyungang, 222001, China. 3. Center of Research Laboratory, Lianyungang Hospital Affiliated to Xuzhou Medical College, 182 Tongguan Road, Lianyungang, 222001, China. 4. Center of Research Laboratory, Lianyungang Hospital Affiliated to Xuzhou Medical College, 182 Tongguan Road, Lianyungang, 222001, China. 18961326009@163.com.
Abstract
PURPOSE: Urinary exosome is an ideal noninvasive, low-cost source of kidney diseases biomarkers. However, many factors effect the isolation of urine-derived exosome. The effect of pH on the yield of exosome isolation under different pH was explored. METHODS: The pH was adjusted for 4, 5, 6, 7 and 8 with hydrochloric acid and sodium hydroxide solution. All samples were incubated for 30 min before differential ultracentrifugation. Exosome-associated protein markers TSG101, ALIX and total concentration of RNA were evaluated by Western-blotting and micro-amount ultraviolet spectrophotometer, respectively. RESULTS: A major loss of urinary exosome was received at pH 8 compared with alkali medium or control group. There was no difference whether or not added EDTA-Na2. CONCLUSIONS: Acidic environment was likely to conducive to the isolation of exosome and maintain its stability and integrity that suggest pH medium needs to be carefully considered and also provide a methodology for future separation exosome.
PURPOSE: Urinary exosome is an ideal noninvasive, low-cost source of kidney diseases biomarkers. However, many factors effect the isolation of urine-derived exosome. The effect of pH on the yield of exosome isolation under different pH was explored. METHODS: The pH was adjusted for 4, 5, 6, 7 and 8 with hydrochloric acid and sodium hydroxide solution. All samples were incubated for 30 min before differential ultracentrifugation. Exosome-associated protein markers TSG101, ALIX and total concentration of RNA were evaluated by Western-blotting and micro-amount ultraviolet spectrophotometer, respectively. RESULTS: A major loss of urinary exosome was received at pH 8 compared with alkali medium or control group. There was no difference whether or not added EDTA-Na2. CONCLUSIONS: Acidic environment was likely to conducive to the isolation of exosome and maintain its stability and integrity that suggest pH medium needs to be carefully considered and also provide a methodology for future separation exosome.
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