(1286-C) Fit-for-Purpose Validation of Cell Signaling Technology® Antibodies for Simple Western™ on the ProteinSimple Jess™.

Abstract: Simple Western™ is a capillary western blotting platform developed by ProteinSimple (a Bio-Techne® brand) that uses capillary electrophoresis to automate protein separation and immunodetection. Simple Western circumvents many of the challenges posed by the manual processes of traditional Western blotting and provides quantitative results in under 3 hours. Moreover, highly specific antibodies used in traditional Western blotting assays are easily adapted to Simple Western systems. Consequently, automated Simple Western platforms are increasingly being used to drive scientific innovation.

Cell Signaling Technology® (CST) is a privately owned and operated producer and supplier of high quality Research-Use-Only (RUO) antibodies, utilizing stringent validation approaches to guarantee exceptional antibody specificity and reproducibility in approved applications. The extensive portfolio of high-quality CST antibodies has played a crucial role in advancing scientific studies using Simple Western platforms. Nearly 500 publications utilizing Simple Western highlight antibodies from CST, representing roughly a quarter of all publications related to Simple Western.

Here, we represent an ongoing collaboration between CST and ProteinSimple to incorporate best-in-class antibodies from CST into ProteinSimple’s automated Simple Western platforms. ProteinSimple and CST jointly developed and implemented a screen to assess CST antibody performance for Simple Western on the ProteinSimple Jess™ system using a 3-point antibody titration across various sample concentrations. Applying this approach, up to three CST antibodies were validated together in the same run using a single 25-capillary cartridge. Altogether, a pilot cohort of 100 CST antibodies was screened in less than two months, achieving an 83% pass rate based on standardized parameters. Our fit-for-purpose validation helps enable faster assay development for Simple Western by providing recommended antibody dilution factors to start optimizing assays.

Currently, over 261 CST antibodies are approved for Simple Western including several unique, modification-state-specific antibodies, with more to come. CST and ProteinSimple maintain a sustained, long‑term partnership for the ongoing validation of antibodies on the Jess platform. Our continuing efforts to validate and guarantee the performance of trusted CST antibodies on Jess de-risks the transition from manual Western blots to automated Simple Western assays.