PURPOSE OF WESTERN BLOT ANALYSIS CODE

It also produces publishable figures which indicate exactly the quantification areas which is extremely important for a transparent measurement. I will try to get in contact with the authors and see if there might be a possibility to link it as plugin into ImageJ (e.g. The method is based on building an antibody:protein complex via specific binding of antibodies to proteins immobilized on a membrane and detecting the bound antibody with one of several detection methods. It is freely available but not completely open source with access to the source code. Western blotting, also known as immunoblotting, is a well-established and widely used technique for the detection and analysis of proteins.

PURPOSE OF WESTERN BLOT ANALYSIS CODE

The design of a quantitative western blot experimentģ.) Since I first tried to code a plugin which would combine the necessary functions for ImageJ, I stumbled over an existing and extremely good tool (also Java) with all the things you would need to run such an analysis (including calibration).Western Blotting Inaccuracies with Unverified Antibodies: Need for a Western Blotting Minimal Reporting Standard (WBMRS).Evaluating Strategies to Normalise Biological Replicates of Western Blot Data.A Defined Methodology for Reliable Quantification of Western Blot Data (Here you see the analysis methos indicated).Quantifying Western blots: Pitfalls of densitometry.So, here a few recommendations and very helpful links (since this is not my invention):ġ.) have a look at the following video seminar from Aldrin Gomes as an introduction to the problem on the experimental side:Ģ.) Here a few publications which help in getting a better understanding before the actial image analysis So, even while often suggested, drawing boxes around a band is not the proper way! 2: The ►Analyze ►Gel tools and the ►Analyze ►Calibrate… function provide theoretically everything what you would need but a clear guideline is still missing. Western Blotting Protocol (Immunoblotting Protocol) Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of proteins using antibodies with fluorescent or chemiluminescent detection. That’s why I posted it here!Įverything I have found in the web so far was mostly going a in the wrong direction leading to non-reliable measurements!

1: Many students try to find a guideline for Western blot “quantification” online while using ImageJ. And I think the topic is important in the aspect of reliability and reproducibility in science. So the following topic is not directly linked to ImageJ necessarily but many of this community might still be interested in. Since once in a while the topic semi-quantitative Western blot analysis pops up and many people in biological sciences who do Western blots or gels are into that, I wanted to share some information on that.

Purpose of western blot analysis