Bio-Rad Laboratories has announced the availability of technical note 5723A, which describes the application of multiplex fluorescent detection that is used to increase western blot throughput
The most common means of analysing protein expression levels is western blotting with detection of a single protein target.
While current methods work well for studying a single target, they are unsuitable for analysing multiple targets, particularly if the target proteins are of unknown or similar size.
To analyse multiple targets, the blot is typically stripped and re-probed.
However, reprobing is a time-consuming process and often some of the target protein on the blot is lost as a result of the stripping procedure.
A recent trend to resolve these issues is the application of fluorescence-based detection of proteins on western blots.
This approach provides the capacity for multiplex analyses as well as greater signal stability relative to chemiluminescent detection.
In addition, simultaneous probing reduces the time and labor required for western blot processing.
Technical note 5723A was designed to help those who are new to fluorescent western blot detection to quickly generate reliable and reproducible results.
Divided into three sections, the first section provides a complete protocol, including materials and their sources.
The Tips section helps assure the success of initial experiments, and the Troubleshooting section helps provide continued success.
