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HTRF Human and Mouse Phospho-α-Synuclein (Ser129) Detection Kit, 500 Assay Points
HTRF Human and Mouse Phospho-α-Synuclein (Ser129) Detection Kit, 500 Assay Points
generic HTRF phospho primary image
| Feature | Specification |
|---|---|
| Application | Cell Signaling |
| Sample Volume | 16 µL |
Product variants
Unit Size: 500 assay points
Part #:
6FSYNPEG
Unit Size: 10,000 assay points
Part #:
6FSYNPEH
Unit Size: 50,000 assay points
Part #:
6FSYNPEY
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).
HTRF Human and Mouse Phospho-α-Synuclein (Ser129) Detection Kit, 500 Assay Points
generic HTRF phospho primary image
HTRF Human and Mouse Phospho-α-Synuclein (Ser129) Detection Kit, 500 Assay Points
Product information
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).
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-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-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.
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.
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.
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.
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
HTRF Human and Mouse Phospho-α-Synuclein (Ser129) Detection Kit, 500 Assay Points
HTRF Human and Mouse Phospho-α-Synuclein (Ser129) Detection Kit, 500 Assay Points
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