Western Blot 2.0 – Sapphire Biomolecular Imager
A quantitative evaluation of western blots, stable signals for years, multiplexing options, and highly sensitive detection – all of this is possible with our Sapphire from Azure Biosystems.
To date, the western blot is the most frequently used technique for detecting proteins. The detection takes place very frequently by means of an enzyme reaction. The chemiluminescence detected in this way is very sensitive and can detect very small amounts of protein, however it does not generate a stable signal. A quantitative statement is also difficult because of the enzymatic reaction.
Fluorescence western blots are suitable for multiplexing and yield stable signals
But especially for scientific publications, a signal must be stable in order to still be detectable even years later, if necessary. To make such a lasting detection possible, an antibody is combined with a fluorophore. The detection of proteins by means of fluorescence has other advantages in addition to the long signal duration.
Detecting several proteins on one and the same blot at the same time is not a problem. The detection with fluorescence is perfectly suited for the use of multiplexing. The possibility of detecting a wide variety of proteins at the same time offers a completely new analysis option: the production of quantitative results!
New possibilities for quantitative western blots
Previously, housekeeping genes were always relied on for the quantitative analysis of western blots, a method which is disputed, however.
Today we are able to produce “true” quantitative western blots. By normalisation on the entire protein (western blot 2.0), it is possible to answer completely new questions. The risk of a false-positive result through a regulated housekeeping gene decreases dramatically.
Chemiluminescence is the most sensitive detection method
Stable signals, multiplexing and quantitative analyses are the greatest advantages of fluorescence detection. But when it comes to sensitivity, the enzymatic detection using chemiluminescence remains undefeated.
This is why we need a detection device which can do both: detection with chemiluminescence and analysis through fluorescence. As a result, it is possible to perform quantitative analyses and highly sensitive research at the same time. Our Sapphire from Azure Biosystems offers precisely this possibility: It makes it possible to generate various detections with one and the same device in high-resolution image quality and at a previously unimagined speed. Scanning with the Azure Sapphire means producing quantitative, reproducible data and, at the same time, using the unique option of several detection methods on the protein level!