Revvity, 63ADK039PEG, HTRF Human Frataxin Detection Kit, 500 Assay Points

Frataxin is a protein localized in the mitochondrion that acts either as an iron chaperone or an iron storage protein. Frataxin mRNA is predominantly expressed in tissues with a high metabolic rate such as the liver, kidneys, adipose tissue, and heart. Reduced expression of Frataxin is the cause of Friedreich's ataxia (FRDA), a lethal neurodegenerative disease. The human Frataxin kit enables the fast monitoring of Frataxin 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 Frataxin is measured using a sandwich immunoassay involving two specific Frataxin antibodies, respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of Frataxin present in the sample. Assay protocol The human Frataxin 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 Frataxin. 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.