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HTRF Human & Mouse Phospho-α-Synuclein (Ser129) Detection Kit, 500 Assay Points

This HTRF kit enables the cell-based detection of phosphorylated alpha-synuclein at Ser129 as a marker of neurodegenerative diseases.

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Feature Specification
Application Cell Signaling
Sample Volume 16 µL

This HTRF kit enables the cell-based detection of phosphorylated alpha-synuclein at Ser129 as a marker of neurodegenerative diseases.

Click to copy promo code to clipboard.
img-icon-save-10-black.svg

March lab savings - buy on Revvity.com or Revvity punchout to SAVE 10% with promo code!

MARCH10

Sale ends 3/24. Terms and conditions apply.

Product Variants
Unit Size: 500 assay points
Part #:
6FSYNPEG
List Price
USD 2,271.53
Your online price:
Unit Size: 10,000 assay points
Part #:
6FSYNPEH
List Price
USD 13,214.42
Your online price:
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).

Overview

The HTRF Phospho-alpha Synuclein (Ser129) cell-based assay kit is ideal for quantifying endogenous phospho-alpha Synuclein phosphorylated on Serine 129.

The alpha Synuclein protein is involved in 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).

Specifications

Application
Cell Signaling
Brand
HTRF
Detection Modality
HTRF
Lysis Buffer Compatibility
Lysis Buffer 1
Lysis Buffer 2
Lysis Buffer 3
Lysis Buffer 4
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
Mouse
Technology
TR-FRET
Therapeutic Area
Neuroscience
Unit Size
500 assay points

Video gallery

Citations

How it works

Phospho-Synuclein (Ser129) assay principle

The Phospho-alpha Synuclein (Ser129) assay measures Synuclein when phosphorylated at Ser129. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer.

The Phospho-Synuclein (Ser129) assay uses 2 labeled antibodies: one with a donor fluorophore, the other with an acceptor. The first antibody was selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both the labeled antibodies, and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the protein’s phosphorylation state under a no-wash assay format.

Phospho how it works phospho alpha synuclein s129 assay principle
Phospho-alpha Synuclein (Ser129) 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 Phospho-alpha Synuclein (Ser129) HTRF detection reagents.

 

This protocol enables the cells' viability and confluence to be monitored.

Phospho how it works phospho alpha synuclein s129 2 plate assay protocol
Phospho-alpha Synuclein (Ser129) 1-plate assay protocol

Detection of Phospho-alpha Synuclein (Ser129) 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.

 

Phospho how it works phospho alpha synuclein s129 1 plate assay protocol

 

Assay validation

Phosphorylation of alpha Synuclein on Serine 129 in Human samples

HEK293 cells were transfected with a plasmid encoding alpha-Synuclein prior to seeding treatment with non phosphorylated fibrils. After cell lysis, different dilutions of lysate were analyzed by HTRF.

Phosphorylation of alpha synuclein was detected in human brain homogenates from patients with Lewy Body Diseases.

assay-validation-alpha-synuclein-phospho-s129-1
assay-validation-alpha-synuclein-phospho-s129-2

Phosphorylation of alpha Synuclein on Serine 129 in Mouse samples

The alpha Synuclein phosphorylation was detected on primary neurons obtained from WT mouse .

Inducible recombinant mouse N2A cells treated with non phosphorylated fibrils displayed high phosphorylation levels of alpha Synuclein.

assay-validation-alpha-synuclein-phospho-s129-3
assay-validation-alpha-synuclein-phospho-s129-4

 

Simplified pathway

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.

phospho-pathway-alpha-synuclein-phospho-s129-kit-6fsynpeg-6fsynpeh-6fsynpey

 

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