Revvity, 64ATA2MPEG, HTRF Human and Mouse Mutant Ataxin 2 Detection Kit, 500 Assay Points

Ataxin 2 is an RNA-binding protein that can be found in different species, organs (predominantly the brain), and cellular locations. In humans, the presence of an extended polyglutamine tract in ataxin 2 is linked to neurodegeneration and contributes to the progression of conditions such as amyotrophic lateral sclerosis (ALS) and spinocerebellar ataxia type 2 (SCA2). Some studies have shown that the polyglutamine expansion correlates with Ataxin 2 protein aggregation. 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
Principle of the HTRF human mutant Ataxin 2 assay The HTRF Mutant Ataxin 2 assay is based on a TR-FRET sandwich immunoassay involving two specific antibodies, one labelled with Eu3+-cryptate (donor) and the other with d2 (acceptor). One antibody is directed against the total Ataxin 2 protein, and the second recognizes specifically the mutant polyQ domain. Both antibodies bind to soluble Mutant Ataxin 2, and the donor-acceptor proximity enables a fluorescent TR-FRET signal. The intensity of the signal is directly proportional to the concentration of soluble Mutant Ataxin 2 present in the sample (cell lysate or tissue lysate). Protocol of the HTRF human mutant Ataxin 2 assay The HTRF Mutant Ataxin 2 assay can be run in a 96- or 384-well low volume white detection plate (20 µL final). As described here, samples (cell/tissue lysates) or standards are dispensed directly into the assay plate for the detection of Mutant HTT by HTRF® reagents. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.