![]() ![]() The proposed method represents an affordable, accurate and reproducible approximation that could be used in the presence of limited resources availability.īackground subtraction Densitometry ImageJ Quantification Western blot.Ĭopyright © 2018 Elsevier B.V. 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. Furthermore, the use of semi-quantitative software as ImageJ (Java-based image-processing and analysis software) is clearly increasing in different scientific fields. 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. In this tutorial we will use ImageJ as a tool to introduce students in the quantitative analysis of virtual bands resembling Western, Northern and Southern. Use the wand and get a quantification value from the intensity of my band. From the generated plots I mark the bell shaped curve with the line tool. To quantify my western blot bands: I need to place a box around each band. Western blot quantification method constitutes a critical step in order to obtain accurate and reproducible results. Hello, I have a question regarding western blot analysis using imageJ. It constitutes a multistep method that allows the detection and/or quantification of proteins from simple to complex protein mixtures. Since its first description, Western blot has been widely used in molecular labs. While uncertain antibody specificity threatens the validity of any Western blot assay 2, 3, 20, 3437, a Western blot performed using an antibody that does not recognise the correct target can only be fixed by replacing it with one that does. ![]()
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