Revvity, 63ADK038PEH, HTRF Human FMRP Detection Kit, 10,000 Assay Points

FMRP (Fragile X Mental Retardation Protein) is a brain protein known to play an important role in normal cognitive development and female reproductive function. fmr1 gene silencing has been correlated to fragile X syndrome onset triggering severe learning deficits or mental retardation. Mutations of the fmr1 gene can also be associated to intellectual disability, premature ovarian failure, autism, Parkinson's disease, developmental delays, and other cognitive deficits. The human FMRP kit enables the fast monitoring of FMRP produced by cells. HTRF assays offer many advantages over other technologies: Homogeneous add-and-read format No wash steps Low background Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates Stable signal, providing flexibility in time of readout or size of assays
Assay principle Human FMRP is measured using a sandwich immunoassay involving two specific anti-FMRP antibodies, respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of the FMRP present in the sample. Assay protocol The human FMRP assay features a two-plate assay protocol, where cells are plated, stimulated, and lysed in the same culture plate. Lysates are then transferred to the assay plate for the detection of Human FMRP. This protocol enables the cells' viability and confluence to be monitored. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay detection can be run in 96- to 384-well plates by simply resizing each addition volume proportionally.