Alpha-Synuclein aggregation Kit
Alpha-Synuclein aggregation detection in cell and brain extracts
This HTRF kit detects cellular alpha Synuclein and can be combined with our phospho-alpha Synuclein S129 kit.
The Total alpha Synuclein cellular assay is designed to monitor the expression level of alpha Synuclein proteins, both phosphorylated and unphosphorylated. It uses the same buffers as our Phospho-alpha Synuclein kit and enables the analysis of phosphorylated and total proteins from a single sample for a better understanding of the alpha Synuclein protein mediated pathogenesis.
The alpha Synuclein protein is involved in the developmnt of synucleinopathies, a group of neurodegenerative diseases that includes Parkinson’s disease (PD), dementia with Lewy bodies (DLB), diffuse Lewy body disease (DLBD), and multiple system atrophy (MSA)
The Total-alpha Synuclein assay quantifies the expression level of Synuclein in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer.
The Total-alpha Synuclein assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of Synuclein in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein’s expression under a no-wash assay format.
The 2 plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before adding Total-Synuclein HTRF detection reagents.
This protocol enables the cells' viability and confluence to be monitored.
Detection of total alpha Synuclein with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required.
This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.
Hela and SH-SY5Y cells were seeded and incubated until confluence at 37°C, 5% CO2.
After cell lysis, the lysates were serially diluted. The total protein content was then quantified using BCA reagent, while alpha Synuclein was quantified with the HTRF Total alpha Synuclein cellular kit.
HTRF total alpha Synuclein does not detect the beta-Synuclein protein.
Alpha-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 alpha synuclein on Ser129 is also associated with a pathological event.
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HTRF and WB compatible guidelines - Technical Notes
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One technology across all samples - Application Notes
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In collaboration with Bayer - Scientific Presentations
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Learn how to reduce time and sample consumption - Application Notes
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