Total alpha Synuclein cellular kit

This HTRF kit detects cellular alpha Synuclein and can be combined with our phospho-alpha Synuclein S129 kit.

Ready-to-use
High sensitivity
Enables fast assay qualification
No-wash

This HTRF kit detects cellular alpha Synuclein and can be combined with our phospho-alpha Synuclein S129 kit.

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Overview

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)

Benefits

SPECIFICITY
PRECISION
DATA NORMALIZATION

Total alpha Synuclein assay principle

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.

Total Synuclein 2-plate assay protocol

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.

Total a Synuclein 2-plate assay protocol

Total alpha Synuclein 1-plate assay protocol

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.

Alpha synuclein level, quantified by HTRF, is more abundant in Hela cells compared to SH-SY5Y

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.

Alpha synuclein level, quantified by HTRF, is more abundant in Hela cells compared to SH-SY5Y

Specifically quantification of the alpha Synuclein, not the beta Synuclein

HTRF total alpha Synuclein does not detect the beta-Synuclein protein.

Parkinson's disease pathway and a-synuclein role

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.

Analysis of the effect of aggregated beta-Amyloid on cellular signaling pathways critical for memory in Alzheimer's disease

Changes in CREB and ERK phosphorylation levels after beta-amyloid treatment - Application Notes

HTRF® phospho-protein platform facilitates the dissection of signaling pathways involved in insulin resistance and metabolic disorders

Multi-tissue cellular modeling and anlysis of insulin signaling - Posters

Simplify complex pathway dissection by combining the power of HTRF® cellular phospho-assays and the flexibility of the CyBi®-FeliX liquid handling system

PI3K/AKT/mTor translational control pathway - Posters

Total alpha Synuclein cellular kit

Overview

Benefits

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