Navarro Paula F, Gil Laura, Fernández-Barredo Salceda
Facultad de Veterinaria y Ciencias Experimentales, Universidad Católica de Valencia San Vicente Mártir, Valencia, Spain.
CEDIVET (Centro Diagnóstico Veterinario), Valencia, Spain.
Bio Protoc. 2022 Aug 5;12(15). doi: 10.21769/BioProtoc.4466.
Capillary electrophoresis (CE) is a laboratory method usually used to separate proteins in body fluids such as serum, cerebrospinal fluid, or urine. Separation of proteins in urine can have clinical applications for evaluating samples from healthy dogs and dogs with proteinuria in a qualitative way, which would not be possible with gel electrophoresis. Other advantages of CE over gel electrophoresis in serum include the reduced separation time (2 min vs. 20 min in a gel), reduction of waste harmful to humans and the environment, and ability to obtain a curve without the need for additional staining. This protocol is divided into four steps. Firstly, urine needs to be prepared prior to dialysis. Secondly, urine needs to undergo dialysis to eliminate compounds that could interfere with separation, and to concentrate the urine. The third step is CE using specific equipment. The last step is to separate the fractions of the phoretograms obtained in the previous step. This method is mostly an automatized process, easily reproducible, and that can be performed in any laboratory, as a part of the diagnostic or follow-up of patients with renal disease. Graphical abstract.
毛细管电泳(CE)是一种实验室方法,通常用于分离血清、脑脊液或尿液等体液中的蛋白质。尿液中蛋白质的分离在临床上可用于定性评估健康犬和蛋白尿犬的样本,而凝胶电泳则无法做到这一点。与血清中的凝胶电泳相比,CE的其他优势包括分离时间缩短(2分钟对比凝胶电泳的20分钟)、减少对人类和环境有害的废物,以及无需额外染色就能获得曲线的能力。本方案分为四个步骤。首先,尿液在透析前需要进行制备。其次,尿液需要进行透析以消除可能干扰分离的化合物,并浓缩尿液。第三步是使用特定设备进行CE。最后一步是分离上一步获得的电泳图谱的各部分。这种方法大多是自动化过程,易于重现,并且作为肾病患者诊断或随访的一部分,可在任何实验室进行。图形摘要。