HTRF Human Phospho-α-Synuclein (Ser129) Aggregation Kit, 500 Assay Points

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HTRF Human Phospho-α-Synuclein (Ser129) Aggregation Kit, 500 Assay Points

This HTRF enables the cell-based quantitative detection of Alpha-Synuclein when phosphorylated at Ser129 and aggregated.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).

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Feature	Specification
Application	Cell Signaling
Sample Volume	16 µL

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This HTRF enables the cell-based quantitative detection of Alpha-Synuclein when phosphorylated at Ser129 and aggregated.

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Product Variants

Unit Size: 500 Assay Points

Part #:

64SYN29AGPEG

List Price

USD 2,147.00

Unit Size: 10,000 Assay Points

Part #:

64SYN29AGPEH

List Price

USD 12,490.00

Product information

Overview

Monitoring the phosphorylation and aggregation states of alpha-synuclein is of great interest for studying the pathogenesis 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 kit provides a solution to rapidly detect and quantify protein aggregates of human alpha-synuclein in cell culture or brain tissue extracts.

Specifications

Other Specifications

Application	Cell Signaling
Brand	HTRF
Detection Modality	HTRF
Lysis Buffer Compatibility	Lysis Buffer 5
Molecular Modification	Phosphorylation
Product Group	Kit
Sample Volume	16 µL
Shipping Conditions	Shipped in Dry Ice
Target Class	Phosphoproteins
Target Species	Human
Technology	TR-FRET
Unit Size	500 Assay Points

Video gallery

How it works

Principle of the HTRF (h) Aggregated Phospho-A-Synuclein Ser129 assay

Alpha-Synuclein aggregates are measured using a sandwich immunoassay involving an anti-a Synuclein monoclonal antibody specific for phospho-Ser129 Alpha-Synuclein aggregates labelled with terbium-Cryptate or d2, ensuring assay quality, reproducibility, and signal quality. The specific HTRF signal generated is proportional to the alpha-Synuclein aggregates.

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Protocol of the HTRF Human Aggregated Phospho-A-Synuclein Ser129 assay

The sample or brain extracts are transferred to the assay plate for the detection of Alpha-Synuclein aggregates. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay detection can be run in 96- to 384-well plates by simply resizing each addition volume proportionally.

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Assay validation

Validation of Aggregated Phospho-A-Synuclein Ser129 detection

A sample generated with aggregated phospho-Alpha-Synuclein (S129) was purified by exclusion phase (G25), resulting in 3 peaks corresponding respectively to oligomers, dimers, and monomers of phospho-Alpha-Synuclein.

Each sample resulting from this purification step was serial-diluted 14 times and plated at 16 µL/well in a low volume white microplate.

2µL of the HTRF d2 detection reagent and 2µL of the HTRF Eu-Tb detection reagent were added, and the signal was recorded after an overnight incubation at RT.

The results show significant detection of the oligomer aggregates only. Closer examinations of the dimer aggregates' detection showed a lower but still detectable signal. Monomers were not detected, demonstrating the specificity of the assay for aggregates only.