Revvity, AL3205F, AlphaLISA Human and Mouse Mutant HTT Detection Kit, 5,000 Assay Points

Overview
Huntingtin (HTT) is a cytoplasmic protein which is highly expressed in the brain, and whose anti-apoptotic role is critical for neuronal survival. The wild-type (WT) protein has a functional structure, with a "normal" polyglutamine (polyQ) domain containing less than 36Q. The mutant HTT protein harbors an abnormally long polyQ tract (> 36Q) which causes the aggregation of the no longer functional protein. HTT aggregation leads to the selective neuronal cell death responsible for Huntington's Disease.
Formats

Our 100 assay point kit allows you to run 100 wells in 96-well format, using a 100 µL reaction volume (10 µL of sample).
Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample

Features

No-wash steps, no separation steps
ELISA alternative technology
Sensitive detection
Broad sample compatibility
Small sample volume
Results in less than 4 hours

AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
How it works
Principle of the AlphaLISA Human & Mouse Mutant HTT assay
The AlphaLISA Mutant HTT assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-target antibody bound to Streptavidin-coated AlphaLISA Donor beads and an anti-target antibody conjugated to AlphaLISA Acceptor beads. One antibody is directed against the total HTT protein, and the second recognizes specifically the mutant polyQ domain. In the presence of the target, both antibodies bind to Mutant HTT, and the beads come into proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules that triggers a cascade of energy transfer within the Acceptor beads, resulting in emission with λmax at 615 nm. The intensity of the signal is directly proportional to the concentration of Mutant HTT present in the sample (cell lysate or tissue lysate).

Protocol of the AlphaLISA human & mouse Mutant HTT assay
The AlphaLISA Mutant HTT assay can be run in a 96- or 384-well detection plate (50 µL final). As described here, samples (cell lysates or tissue lysates) or control lysate are dispensed directly into the assay plate for the detection of Mutant HTT by AlphaLISA reagents. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
Application-Protein Quantification
Automation Compatible-Yes
Brand-AlphaLISA
Detection Modality-Alpha
Product Group-Kit
Protocol Time-3.5h at RT
Sample Volume-5 µL
Shipping Conditions-Shipped in Blue Ice
Target-HTT
Target Class-Biomarkers
Target Species-Human Mouse
Technology-Alpha
Therapeutic Area-Neuroscience
Unit Size-5,000 assay points