Description
α-Synuclein is a neuronal protein encoded by the SNCA gene that regulates synaptic vesicle trafficking and subsequent neurotransmitter release. α-Synuclein is a prominent component of intracellular fibrillary aggregates in the brains of patients suffering from synucleinopathies. This protein represents a key molecular hallmark for Parkinson’s disease, dementia with Lewy bodies, or multiple system atrophy. The extensive phosphorylation of α-Synuclein on Ser129 is also associated with a pathological event.
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.
In the AlphaLISA™ SureFire® Ultra™ assay, Donor beads are coated with streptavidin to capture one of the antibodies, which is biotinylated. Acceptor beads are coated with a proprietary CaptSure™ agent that immobilizes the other antibody, labeled with a CaptSure™ tag. In the presence of target protein, the two antibodies bring the Donor and Acceptor beads close together, generating signal. The amount of light emission is directly proportional to the amount of phosphoprotein present in the sample.
AlphaLISA™ SureFire® Ultra™ kits are compatible with:
Cell and tissue lysates
Antibody modulators
Biotherapeutic antibodies
Alpha SureFire® kits can be used for:
Detecting alpha synuclein aggregation
Cellular kinase assays
Receptor activation studies
Screening
Replacing an alpha synuclein ELISA kit
AlphaLISA SureFire Ultra protein dimer / aggregate detection assay principle
The AlphaLISA SureFire Ultra dimer/aggregate assay measures the levels of a given protein in a dimeric, multimeric or any other aggregated form in a biological sample (e.g. cell lysate).
The assay uses a single antibody clone tagged with both CaptSure™ peptide and biotin, that recognizes exposed epitopes on the multivalent target protein. 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 a CaptSure tag. Donor Beads are coated with streptavidin to capture the same specific antibody, which is biotinylated. In the presence of the multivalent protein, the two antibodies bring the Donor and Acceptor Beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor Bead, allowing for the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of multimeric protein present in the sample.
AlphaLISA SureFire Ultra dimer / aggregate detection 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 dimer or aggregate AlphaLISA SureFire Ultra detection reagents. This protocol enables cell viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.
AlphaLISA SureFire Ultra protein dimer / aggregate detection one-plate assay protocol
Detection of the dimer or aggregate 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.
