Revvity, 63ADK012PEH, HTRF Human MBNL1 Detection Kit, 10,000 Assay Points

MBNL1 is involved in RNA-protein complexes with MBNL2 and DMPKRNA. In myotonic dystrophy 1 (DM1), known as a progressive neuromuscular disorder, aggregation of the mutant DMPKRNA with protein complexes has been associated to aberrant splicing of the insulin receptor. MBNL1 has been shown to bind CUG repeat sequences on RNA sequences. 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 MBNL1 is measured using a sandwich immunoassay involving two specific anti-MBNL1 antibodies respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of human MBNL1 present in the sample. Assay protocol The MBNL1 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 MBNL1. 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.