Reprobe Your Immunoassay Samples Using RePlex With Jess

The identification and quantification of protein expression levels is critical to understanding biological processes like those that drive cancer and other diseases. Unfortunately, limited samples sizes are far too common in these research areas, leaving researchers desiring more data from their limited samples. To measure protein expression by traditional methods like the Western blot, researchers must sacrifice most, if not all, of their sample for only a few data points, which are poorly reproducible and semi-quantitative at best. To overcome these limitations, Simple Western™ with Jess™ is an automated, capillary-based immunoassay that provides highly reproducible and quantitative data with a tiny amount of material. With just 3 μL of sample, Jess offers pg-level sensitivity on up to 25 samples in 3 hours, all in a fully automated fashion. Now, Jess comes with RePlex™ so that you can reuse the same 3 μL of sample for automated sequential rounds of probing.

In this application note, we show how you can use RePlex to detect and quantify pan and phospho isoforms of proteins such as those in the PI3K/Akt signaling pathway. This pathway modulates cell growth, survival, and apoptosis, and it is frequently altered in human cancers, contributing resistance to radiation and chemotherapy treatment.