BMG LABTECH recently posted a new application note (#255) that describes the use of the Leading LightTM Sclerostin-LRP Screening System from EnzoLife Sciences which enables the detection of Wnt-pathway modulators.
This application is important in the study of Wnt effects on bone formation which have been characterized to involve the binding to receptors on bone cells leading to regulation of transcription that promotes bone formation. Specifically, Wnt binds to the LRP 5/6 receptor and the effect of Wnt can be atagonized by Sclerostin when it interacts with the LRP 5/6 receptor leading to inhibition of bone formation. Enzo has exploited the LRP-Sclerostin interaction to produce a system that can identify compounds which block this interaction and could serve as treatments for osteoporosis.
Sclerostin-LRP System Assay Principle
In the Leading LightTM Sclerostin-LRP Interaction System, 96 well plates are coated with Sclerostin and binding of LRP can be seen since LRP5 is linked to the enzyme alkaline phosphatase (AP). After washing steps an AP substrate is added so that LRP5 which remains bound to the Sclerostin on the plates will be revealed by a chemiluminescent signal. Treatments which alter the interaction between Sclerostin and LRP5 can thus be identified based on changes in the luminescent signal which can be detected.
In this application note we found that detection of this chemiluminescent signal can be accomplished in a highly sensitive manner using the LVF-monochromator available on the CLARIOstar®! Using the CLARIOstar® the emission profile for the chemiluminescent signal can be observed. Furthermore, the unique ability of the CLARIOstar® to employ bandpasses up to 100 nm is displayed!