Simple Western Sized-Based Assay Data
Viewing Size-Based Assay Data
Simple Western size-based assay data is processed automatically in Compass for Simple Western™. Sample data is displayed by lane in a virtual blot-like image similar to traditional results with one big exception—not only do you get more information, but you also get it as soon as the assay is complete.
Quantitative results like signal height, area, % area, width, S/N, and baseline values for each immunodetected protein are presented in the results table automatically, which may be exported for further analysis if desired.
The lanes in this view can be rearranged, eliminating the need to splice lanes together with 3rd party software or to rerun samples so that they are adjacent to each other.
If you're more familiar with capillary electrophoresis, you can see your results in graph view with a more traditional electropherogram view too. This Cartesian representation of immunoassay data provides quantitative information for signal intensity, which is derived from the area under the curve. Like the lane view above, Compass for Simple Western automatically calculates signal height, area, % area, width, S/N, and baseline values, and may be exported to your favorite statistical analysis software.
Detectable Molecular Weight Range
With kits offering sizing ranges from 2 kDa to 440 kDa, Simple Western assays support the detection of varying protein sizes, resolving small polypeptides or large target proteins like dystrophin.
Simple Western offers Separation Modules of the following three molecular weight ranges:
- 2–40 kDa
- 12–230 kDa
- 66–440 kDa
The Simple Western assay data image shows a panel of proteins run using the 12–230 kDa (left) and a 66–440 kDa (right) Separation Modules.
Reproducible, Quantitative Size-Based Assay Data
Our Simple Western systems give you the detection and quantitation you need, whether you’re looking for absolute or relative quantitation or wanting to simultaneously quantitate high and low protein concentrations.
Simple Western assays are fully automated by Jess, Abby, Peggy Sue, or Sally Sue and easily standardized, making your results much more reproducible. And there’s no blotting step, so inconsistencies caused by protein transfer are eliminated, which really improves your quantitation too!
Figure (right): Phosphoinositide 3-kinase (PI3K) expression in an MCF-7 lysate. Ten replicates of MCF7 lysate at 1 µg/µL were analyzed using a PI3K-specific primary antibody. The predicted molecular weight of PI3K is 110 kDa. Simple Western assay data reported an average molecular weight of 107 kDa with a 0.5% CV for sizing and an 8.7% CV for signal intensity (area).
Dynamic Range Assay
Simple Western provides a linear dynamic range assay of approximately 3 orders of magnitude, letting you detect proteins over a wider concentration range, not to mention with more accurate quantitation than traditional Western blot.
Customer Success Stories
Professor and Chair, University of Birmingham, UK
Postdoctoral Researcher, CSU Fullerton