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自动化与传统蛋白质免疫印迹法的比较

Comparison of Automated and Traditional Western Blotting Methods.

作者信息

Sormunen Aino, Koivulehto Emma, Alitalo Kari, Saksela Kalle, Laham-Karam Nihay, Ylä-Herttuala Seppo

机构信息

A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, FI-70211 Kuopio, Finland.

Translational Cancer Biology, University of Helsinki, FI-00014 Helsinki, Finland.

出版信息

Methods Protoc. 2023 Apr 20;6(2):43. doi: 10.3390/mps6020043.

Abstract

Traditional Western blotting is one of the most used analytical techniques in biological research. However, it can be time-consuming and suffer from a lack of reproducibility. Consequently, devices with different degrees of automation have been developed. These include semi-automated techniques and fully automated devices that replicate all stages downstream of the sample preparation, including sample size separation, immunoblotting, imaging, and analysis. We directly compared traditional Western blotting with two different automated systems, iBind™ Flex, which is a semi-automated system designed to perform the immunoblotting, and JESS Simple Western™, a fully automated and capillary-based system performing all steps downstream of sample preparation and loading, including imaging and image analysis. We found that a fully automated system can save time and importantly offer valuable sensitivity. This is particularly beneficial for limited sample amounts. The downside of automation is the cost of devices and reagents. Nevertheless, automation can be a good option to increase output and facilitate sensitive protein analyses.

摘要

传统的蛋白质免疫印迹法是生物学研究中最常用的分析技术之一。然而,它可能耗时且缺乏可重复性。因此,人们开发了不同自动化程度的设备。这些设备包括半自动化技术和全自动化设备,后者可复制样品制备下游的所有步骤,包括样品大小分离、免疫印迹、成像和分析。我们直接将传统蛋白质免疫印迹法与两种不同的自动化系统进行了比较,即iBind™ Flex(一种设计用于进行免疫印迹的半自动化系统)和JESS Simple Western™(一种全自动化的基于毛细管的系统,可执行样品制备和上样下游的所有步骤,包括成像和图像分析)。我们发现,全自动化系统可以节省时间,并且重要的是能提供有价值的灵敏度。这对于有限的样品量尤其有益。自动化的缺点是设备和试剂的成本。尽管如此,自动化仍是提高产量和促进灵敏蛋白质分析的一个不错选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01e1/10142486/77776e270520/mps-06-00043-g001.jpg

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