ImageJ Western Blot Quantification: A Cost-Effective Solution for Accurate Results

Western blot analysis is a powerful tool for protein detection, allowing researchers to detect and/or quantify proteins from simple to complex protein mixtures. However, the process can be time-consuming and requires technical expertise. In this article, we will explore how ImageJ software, coupled with an office scanner, can be used for accurate Western blot quantification at an affordable cost.

Background subtraction is a crucial step in Western blot analysis, as it helps to eliminate non-specific binding and background noise. Our proposed method involves using a new image background subtraction technique, which has been shown to provide accurate results even with limited resources availability.

In our study, we used an office scanner to scan the Western blot membrane and visualized it in the 800 channel. The resulting image was then analyzed using ImageJ software, which is a free NIH software called Fiji or ImageJ. This software is widely used in different scientific fields due to its ease of use and flexibility.

The quantification process involves several steps, including background subtraction, thresholding, and particle analysis. By using these techniques, researchers can accurately quantify the amount of protein present in their samples. In our study, we successfully quantified phosphorylated Ser139 on histone variant H2A.X (H2AX) stained images of tissue or cells.

The benefits of using ImageJ for Western blot quantification are numerous. For one, it is a cost-effective solution that eliminates the need for expensive dedicated equipment. Additionally, it provides accurate and reproducible results, making it an ideal choice for researchers with limited resources availability.

In conclusion, our proposed method represents an affordable, accurate, and reproducible approximation that can be used in the presence of limited resources availability. By using ImageJ software and an office scanner, researchers can easily quantify Western blots and obtain accurate results without breaking the bank.

Imagej western blot quantification