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The dangers of overexposing western blots

It’s like, how much more black could this be? And the answer is none. None more black.

Nigel Tufnel, lead guitarist of Spinal Tap.


None more black (or none more white) is a real concern when you’re talking about imaging western blots. If you only want to verify the presence of a protein, you can stop reading here, because even an overexposed image will tell you that there is some protein present in your sample. But an overexposed image of a western blot is essentially useless when you’re … Continue Reading

Analyzing western blots with Image Studio Lite

Image Studio Lite is a free software package from LI-COR Biosciences aimed at life scientists that want to analyze gels, western blots, dot blots, and other similar lab outputs. Although the Lite version is free, there is a more comprehensive paid version of the software that aims to easily integrate with the apparatus that Licor also sells. However, the free Image Studio Lite is more than sufficient for basic analysis of western blots, as I’ll show below.

Obviously you need to start by downloading and installing Image Studio Lite. The first time … Continue Reading

Analyzing gels and western blots with ImageJ

The following information is an updated version of a method for using ImageJ to analyze western blots from a now-deprecated older page. If you need a peer-reviewed citation for the methods outlined below, you may cite  Stael, S., L.P. Miller, Á.D. Fernández-Fernández and F. Van Breusegem (2022). Detection of Damage-Activated Metacaspase Activity by Western Blot in Plants. Plant Proteases and Plant Cell Death: Methods and Protocols. M. Klemencic, S. Stael and P.F. Huesgen. New York, NY, Springer US: 127-137 https://doi.org/10.1007/978-1-0716-2079-3_11

If you’re looking for a more comprehensive workflow option for your western blot … Continue Reading