Label Independent, Inc. is a U.S.-based contract lab providing label-free biochemical and cell-based assays. Call Us Today at 866-617-DATA to discuss your assay.
Label-Free cell-based assays provide data based on actual cellular response to various agonists. These assays are of particular importance in GPCR studies where the traditional methods rely on downstream effects such as calcium mobilization.
In traditional assay, GPCR agonists are detected by their ability to create a downstream Ca++ mobilization event. These events are tracked using fluorescent dyes which increase in fluorescent intensity as local calcium levels increase. This may be valuable for Gq-coupled receptors. In cases where Gi/Gs coupled receptors are involved, cAMP assays are more appropiate. These methods require the use of engineered cells which over-express the receptor of interest. Furthermore, these cells must be incubated with various dyes, require washing steps, and are sensitive to autofluorescent compounds given that the method of detection is fluorescent intensity.
Label-Free GPCR assays offer several key advantages. Since the method of detection relies on actual cellular events including protein trafficking and cytoskeletal rearrangement, the assay is more representative of the endogenous state. Furthermore, cells do not have to engineered. Cells can display endogenous levels of the receptor of interest (as few 2000-4000 copies/ cell).
In a single Label-Free assay it is possible to determine of the agonist acts on Gi, Gq, Gs signal transduction pathway. Furthermore, time-resolved signalling allows for the determination of the actual pathway involved. Therefore, in a single label-free assay it is possible to determine if a compound ellicits a cellular event, and more importantly the optical signature can determine if the response is Gi, Gq, or Gs mediated.
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