How Does Jess Work?
Simple Western assays are automated, capillary-based immunoassays that solve many of the challenges that come with traditional Westerns. All assay steps from protein separation, immunoprobing, detection, and analysis are fully automated. The variability in the manual processes that impact reproducibility, quantitation, time to result, and overall data reliability is eliminated. Best of all, the transition is simple—the same antibodies you use for Western blotting protocols can be used with Simple Western assays.
Your sample, separation matrix, stacking matrix, antibodies, and reagents are loaded automatically from a specially designed plate. Jess begins by aspirating the separation matrix and then the stacking matrix into each capillary. Next, your sample is loaded, and capillaries are lowered to make contact with running buffer. Voltage is applied to enable separation by molecular weight. Once the separation is complete, UV light immobilizes the proteins to the capillary wall. With proteins now immobilized and the matrix cleared of the capillary, Jess starts the immunoprobing process. In just 3 hours you'll have quantitative, size-based data ready for analysis.
Jess gives you four different ways to analyze proteins:
- Multiplex detection: Maximize your time and sample, and get the information you need in one shot by multiplexing in chemiluminescence and infrared (IR) and near-infrared (NIR) fluorescence channels. The Stellar™ NIR/IR Modules bring industry-leading fluorescence detection sensitivity for Western workflows. Take multiplexing even further with Superplex to assay in chemiluminescence and 2-color fluorescence with Protein Normalization at the same time.
- RePlex™: Why bother stripping and reprobing? RePlex lets you run two immunoassays within the same capillary to get all your rich protein characterization data from just one sample!
- Protein Normalization: Jess gives you an easy way to see if your samples contain a consistent protein load—just load the Protein Normalization reagent into the assay plate and Jess will take care of the rest. Jess's proprietary fluorescent reagent measures proteins immobilized in the same capillary as your immunoassay. You can effectively normalize expression of your target protein to get accurate and consistent data, giving you confidence in your results. And with Superplex you can assay in chemiluminescence and 2-color fluorescence on top of Protein Normalization.
- Blot imaging: Still doing traditional Westerns? You can image chemiluminescence Western blot with Jess's built-in blot imaging system.
What does the data look like? At the end of the run, Simple Western data are processed automatically in Compass for Simple Western. Simple Western data are displayed by lane in a virtual blot-like image similar to traditional Western blot results. If you're more familiar with capillary electrophoresis, you can see your results in a more traditional electropherogram view too. Quantitative results such as molecular weight, signal intensity (area), % area, and signal-to-noise for each immunodetected protein are presented in the results table automatically.