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Urine Proteomics in Kidney Disease Biomarker Discovery by Youhe Gao

By Youhe Gao

This booklet systematically summarizes the tips and applied sciences utilized in urine proteome research. It argues that adjust is the middle of biomarker definition because the physique makes use of its homeostatic mechanisms to right alterations within the blood. which means urine is perhaps a greater resource of biomarkers than blood. A roadmap to the urinary biomarker period is proposed, and researchers are reminded of the capability possibilities and dangers of their learn layout. Kidney illnesses are emphasised as they produce the main major adjustments in urine. This publication attempts to teach researchers and graduate scholars, who're in or getting into the sphere, "all issues thought of" instead of "the present affair".

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X 14. Thongboonkerd V, Chutipongtanate S, Kanlaya R (2006) Systematic evaluation of sample preparation methods for gel-based human urinary proteomics: quantity, quality, and variability. J Proteome Res 5(1):183–191. 1021/pr0502525 15. Lifshitz E, Kramer L (2000) Outpatient urine culture: does collection technique matter? Arch Intern Med 160(16):2537–2540 16. Schaub S, Wilkins J, Weiler T, Sangster K, Rush D, Nickerson P (2004) Urine protein profiling with surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry.

To demonstrate the orientation of proteins within the low-density urinary vesicles, the authors used immunogold electron microscopy and antibodies against epitopes on the cytoplasmic side of the integral membrane proteins AQP2 and NCC, or antibodies against epitopes on the external side of aminopeptidase N and CD9. Analogous to exosomes found in other bodily fluids, characteristic of vesicles found 5 Exosomes in Urine Biomarker Discovery 45 Fig. 1 Negative staining of urinary exosomes from normal human subjects using electron microscopy at 41,000× magnification.

1 ml elution buffer/cm2 membrane was added. The membrane in the elution buffer was mixed well by vortexing for 10 min at room temperature and then by ultrasound for 15 min in an ultrasonic cleaner at room temperature. The supernatant was collected by spinning down the membrane. The protein was concentrated using a centrifugal filter with a molecular weight cutoff of 3,000 Da or by protein precipitation with 26 L. Liu et al. Fig. 1 Comparing the urinary proteins after removing cells in urine by centrifugation or filtration, respectively [1] chloroform/methanol (if the detergent needed to be removed for the downstream analysis such as protein quantification and LC-MS/MS analysis).

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