Software Choice Can Reduce Western Blot Data Analysis Variability

Ways to Reduce Western Blot Data Analysis Variability

Friday, 28 February, 2020


empiria studio,western blot analysis software,image studio

After you have run a quantitative Western blot, there are several steps to take in your analysis to prepare for publication. It is important to keep in mind that data quantification and analysis should be conducted in a way that does not add variability or error to your results.

Analysis and quantification software can help you determine the difference in band intensity between target bands. While there are many software options to choose from, not all of them will provide accurate data analysis. Software can vary widely in their capabilities and analysis methods, and some can even manipulate and change your data.

Figure 1. (A-B) Two brightness and contrast settings were applied across an image in ImageJ. (C) The normalized signal per lane was graphed for each setting.
Figure 2. (A-B) Two brightness and contrast settings were applied across an image in Image Studio. (C) The normalized signal per lane was graphed for each setting.

Image Studio™ Software

When analyzing Western blots, look for a software that does not change the raw data and adjusts the image display uniformly. You should be able to apply brightness, contrast, and color balance adjustments without altering your data. Additionally, a software should subtract local background and export your data for statistical analysis or graphical comparisons. Two common software packages for Western blot quantification and analysis are Image Studio Software and Empiria Studio Software.

Journals require that your images were not manipulated and that raw data were not compromised in a way that positively or negatively affects your conclusions. As a quantification software, Image Studio does not modify images, so your results are not jeopardized. It saves your data and image display adjustments, so when it is time to publish, you know precisely what you did to get those numbers. Additionally, Image Studio Software makes it easy to sort through images and quickly quantify your bands.

Empiria Studio® Software

Empiria Studio is designed to provide you reliable answers during each analytical step. As an analysis software that incorporates publisher recommendations, Empiria Studio creates a strong foundation for producing high quality quantitative Western blots. It allows you to analyze these blots, determine target expression, compare replicates, conduct statistical analysis, and share the results all within easy-to-follow workflows. These steps can help you mitigate experimental issues by validating your antibodies and housekeeping protein and determining the combined linear range of your targets.

Additionally, its patent-pending Adaptive Background Subtraction (ABS) works automatically and specifically adapts to each image for clearer data. Empiria Studio also allows you to view your results, draw conclusions, and share findings – all in one place.

Looking for more ways to reduce variability in your data analysis? Review our Best Practices for Quantitative Western Blot Data Analysis outline for helpful suggestions.

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