Stripping & reprobing traditional Western blots is often attempted to gain more information from a single blot, however, it can lead to unpredictable protein losses and high signal variability which makes obtaining quantitative data nearly impossible. The RePlex™ feature in Simple Western™ is an optimized and automated two-step immunoassay that enables quantitative, multiplexed protein expression data without loss in signal intensities and without affecting epitope binding.
In this webinar, Dr. Lorrain will describe how RePlex assays on Simple Western have been utilized at the Protein Analysis Facility at the University of Lausanne to advance three specific, multiplexed bioassays. Specifically, she will describe the design & optimization of an assay where two antibodies from the same species but requiring different detection channels were utilized as well as studies comparing several common housekeeping proteins with total protein normalization. In sum, the results of these studies provide guidance on how to carefully design RePlex experiments, especially when dealing with strong chemiluminescence signals in the first step and highlight the flexibility in assay design, multiplexing ability, and accurate quantitation afforded by RePlex sequential immunoassays on Simple Western.
Learn more about how RePlex provides the ability to:
- Quantify expressed phosphorylated target and total target levels.
- Normalize your data with total protein detection in the same capillary.
- Greater flexibility in immunoprobe selection.
- More data points per sample.
- Saving time and money on consumables.
||Séverine Lorrain, PhD
Senior Technician, Protein Analysis Facility
University of Lausanne
Dr. Séverine Lorrain is a molecular biologist that enjoyed deciphering the signaling pathways involved in plant interactions and responses to their environment during her Ph.D. and post-doctoral studies. After a short stay at the Lausanne genomic facility, she moved to the protein analysis facility in the same university to be in charge of antibodies-based detection platforms. She first set-up the Reverse Phase Protein Array platform before mainly focusing on the Simple Western Jess technology.
||Chris Heger, PhD
Director of Applications Science
Analytical Solutions Division of Bio-Techne
Dr. Chris Heger, Director of Applications Science for the Analytical Solutions Division of Bio-Techne, received his doctoral degree in Pharmacology from Cornell University and completed his post-doctoral training at the National Cancer Institute in Antibody Development, Purification and Technology Evaluation. Chris joined Protein Simple in 2011, starting as a Field Applications Scientist, and subsequently further building the East Coast support team as Field Manager. During these years, Chris a developed broad but deep understanding of the instrumentation and their application across diverse research areas. In 2016, Chris moved to California to lead the Applications Science group, chartered with creating scientific collateral, fostering collaborations, training, and application development. Chris is an expert in immunoassays, capillary electrophoresis, protein and antibody purification, and chromatography.