PURECLTM HRP substrates - Western Blotting for advanced users
Sensitivity is one of the most widely advertised properties of chemiluminescent substrates. Traditionally, a substrate is considered to be more sensitive if it enables shorter measuring times or leads to a stronger signal in a given time. According to the logic: a faster substrate will automatically visualize weaker signals better.
Problem: in fact, sensitivity is confused with reactivity.
In order to be able to detect a specific antigen (protein) in low concentration, it is imperative that the background is avoided in the experiment if possible. Some of the supposedly most sensitive (or most reactive) substrates on the market - however, are known for the unwanted background development they promote. Conversely, rather background-neutral substrates are often not very sensitive.
Sensitivity and specificity are therefore two sides of the same medal.
In chemiluminescence western blots, numerous factors are responsible for the formation of the background. These include, for example: the affinity of the antibody, the efficiency of the blocking process or the quality of the measuring device - to name just a few. When choosing the substrate, however, its own influence on the background formation is often underestimated.
This is exactly where our PURECLTM HRP substrates come into play.
They have been optimized on FUSION imagers with special regard to background neutrality. A key aspect of FUSION technology is the particularly low-noise measurement.Both substrates are therefore suitable for the sensitive detection of Western or dot blot, but are not limited to FUSION imagers.
PURECLTM Ultra HRP substrate is extremely reactive and enables very short process times with maximum sensitivity. It was originally developed for the shortened protocol of the PURITYTM HRP detection system (Western Blot 2.0) but is also suitable for the classic sequential method.
PURECLTM Dura HRP substrate is slightly less reactive, but has a signal duration of up to several hours. Due to its enormous experimental robustness, PURECL Dura is suitable for a very wide range of different applications:From the first attempts with new antibodies to the low-expressed phosphoproteins. Even the high-throughput laboratory, which sometimes uses substrate several times for economic reasons, appreciates PURECLTM Dura as the perfect all-rounder.