Gallo-Oller Gabriel, Ordoñez Raquel, Dotor Javier
Childhood Cancer Research Unit, Department of Women's and Children's Health, Karolinska Institutet, Astrid Lindgren Children's Hospital, Stockholm, Sweden.
Area Oncohematología, Centro de Investigación Médica Aplicada (CIMA), Pamplona, Navarra, Spain.
J Immunol Methods. 2018 Jun;457:1-5. doi: 10.1016/j.jim.2018.03.004. Epub 2018 Mar 6.
Since its first description, Western blot has been widely used in molecular labs. It constitutes a multistep method that allows the detection and/or quantification of proteins from simple to complex protein mixtures. Western blot quantification method constitutes a critical step in order to obtain accurate and reproducible results. Due to the technical knowledge required for densitometry analysis together with the resources availability, standard office scanners are often used for the imaging acquisition of developed Western blot films. Furthermore, the use of semi-quantitative software as ImageJ (Java-based image-processing and analysis software) is clearly increasing in different scientific fields. In this work, we describe the use of office scanner coupled with the ImageJ software together with a new image background subtraction method for accurate Western blot quantification. The proposed method represents an affordable, accurate and reproducible approximation that could be used in the presence of limited resources availability.
自首次被描述以来，蛋白质免疫印迹法（Western blot）已在分子实验室中广泛应用。它是一种多步骤方法，可用于检测和/或定量从简单到复杂的蛋白质混合物中的蛋白质。蛋白质免疫印迹法定量方法是获得准确且可重复结果的关键步骤。由于光密度分析所需的技术知识以及资源可用性，标准办公扫描仪常被用于已显影的蛋白质免疫印迹膜的成像采集。此外，在不同科学领域，像ImageJ（基于Java的图像处理和分析软件）这样的半定量软件的使用明显在增加。在这项工作中，我们描述了将办公扫描仪与ImageJ软件结合使用，并采用一种新的图像背景扣除方法来进行准确的蛋白质免疫印迹法定量。所提出的方法是一种经济实惠、准确且可重复的方法，在资源有限的情况下也可使用。
