Description
Overview
Alpha-Synuclein (α-Syn) is a small, intrinsically disordered protein predominantly expressed in neurons, particularly in presynaptic terminals. It plays a crucial role in synaptic function, vesicle trafficking, neurotransmitter release and synaptic plasticity. In its normal state, α-Syn exists as a soluble monomer, but it can misfold and aggregate into oligomers, fibrils, and Lewy bodies. These aggregates are a hallmark of synucleinopathies, including Parkinson's disease (PD), Dementia with Lewy Bodies (DLB), and Multiple System Atrophy (MSA). Misfolded α-Syn disrupts cellular homeostasis, impairs synaptic function, and propagates toxic fibrils between neurons in a prion-like manner, leading to neurodegeneration. Therapeutic strategies aim to reduce α-Syn aggregation, enhance its degradation, and prevent its pathological spread.
The AlphaLISA
SureFire Ultra
Human and Mouse Phospho-α-Synuclein (Ser129) Aggregate Detection Kit is a sandwich immunoassay for the quantitative detection of phospho-α-Synuclein (Ser129) aggregate in cellular lysates, using Alpha Technology.
Formats:
The HV (high volume) kit contains reagents to run 100 wells in 96-well format, using a 60 μL reaction volume.
The 500-point kit contains enough reagents to run 500 wells in 384-well format, using a 20 μL reaction volume.
The 10,000-point kit contains enough reagents to run 10,000 wells in 384-well format, using a 20 μL reaction volume.
The 50,000-point kit contains enough reagents to run 50,000 wells in 384-well format, using a 20 μL reaction volume.
AlphaLISA
SureFire Ultra
kits are compatible with:
Cell and tissue lysates
Antibody modulators
Biotherapeutic antibodies
AlphaLISA
SureFire Ultra
kits can be used for:
Cellular kinase assays
Receptor activation studies
High-throughput screening for preclinical studies
How it works
Phospho Aggregation-AlphaLISA
SureFire Ultra
assay principle
The Phospho Aggregation-AlphaLISA
SureFire Ultra
assay measures the level of phosphorylated aggregates of a protein target in a cell lysate. This assay uses a single phospho-specific antibody that recognizes an exposed phospho-epitope on the aggregated target protein, that is conjugated with either a CaptSure™ tag or with Biotin. AlphaLISA assays require two bead types: Acceptor and Donor beads.
Acceptor beads are coated with a proprietary CaptSure agent to specifically immobilize the antibody labeled with the CaptSure tag. Donor beads are coated with Streptavidin to capture the biotinylated antibody. In the presence of phosphorylated aggregate protein, the Donor and Acceptor beads are brought into close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, allowing the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of phosphorylated aggregate protein present in the sample.
Phospho Aggregation-AlphaLISA
SureFire Ultra
two-plate assay protocol
The two-plate protocol involves culturing and treating the cells in a 96-well plate before lysis, then transferring lysates into a 384-well Optiplate plate before the addition of Phospho Aggregation-AlphaLISA
SureFire Ultra
detection reagents. This protocol allows for the cells viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.
Phospho Aggregation-AlphaLISA
SureFire Ultra
one-plate assay protocol
Detection of aggregated target protein with AlphaLISA
SureFire Ultra
reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol allows for miniaturization while maintaining robust AlphaLISA
SureFire Ultra
quality.
