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

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  • Ready-to-use Ready-to-use
  • High sensitivity High sensitivity
  • Enables fast assay qualification Enables fast assay qualification
  • No-wash 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 a Synuclein assay principle

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.

Total-TBK1 1-plate assay protocol

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.

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.

Parkinson's disease pathway
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

Detection of human tau protein aggregation

For researcher working on PD or AD - Application Notes

Cisbio lysis buffer compatibility

Cell Signaling: Biomarkers, Phospho- & total-protein Assays - Flyers

Species compatibility

Cell Signaling: Biomarkers, Phospho- & total-protein assays - Flyers

HTRF Alpha-tubulin Housekeeping kit

Properly interpret your compound effect - Application Notes

HTRF cell-based phospho-protein data normalization

Valuable guidelines for efficiently analyzing and interpreting results - Application Notes

Optimize your HTRF cell signaling assays on tissues

HTRF and WB compatible guidelines - Technical Notes

Unleash the potential of your phosphorylation research with HTRF

A fun video introducing you to phosphorylation assays with HTRF - Videos

How to run a cell based phospho HTRF assay

3' video to set up your Phospho assay - Videos

Key guidelines to successful cell signaling experiments

Mastering the art of cell signaling assays optimization - Guides

Exploring LRRK2 phosphorylation with HTRF® assays

LRRK2 kit, a game changer in neuroscience - Posters

Exploring Synuclein phosphorylation and aggegation with HTRF assays

Alpha Synuclein kits, perfect tools to study synucleinopathy - Posters

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Best practices for analyzing brain samples with HTRF® phospho assays for neurosciences

Insider Tips for successful sample treatment - Technical Notes

Product Insert a-Syn total Kit / 6FNSYPEG-6FNSYPEH

6FNSYPEG-6FNSYPEH - Product Insert

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Best practices for analyzing tumor xenografts with HTRF phospho assays

Protocol for tumor xenograft analysis with HTRF - Technical Notes

HTRF® cell signaling platform combined with iCell® Hepatocytes

A solution for phospho-protein analysis in metabolic disorders - Posters

HTRF phospho-assays reveal subtle drug-induced effects

Detailed protocol and direct comparison with WB - Posters

Universal HTRF® phospho-protein platform: from 2D, 3D, primary cells to patient derived tumor cells

Analysis of a large panel of diverse biological samples and cellular models - Posters

HTRF phospho assays reveal subtle drug induced effects in tumor-xenografts

Tumor xenograft analysis: HTRF versus Western blot - Application Notes

HTRF phospho-total lysis buffer: a universal alternative to RIPA lysis buffers

Increased flexibility of phospho-assays - Application Notes

Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling

Analyse of PI3K/AKT/mTor translational control pathway - Application Notes

Side-by-side comparison of HTRF, Western Blot, ELISA and AlphaScreen® SureFire®

Do all cell-based kinase assays perform similarly? - Posters

How to run a cell based phospho HTRF assay

What to expect at the bench - Videos

Molecular basis of neuroinflammation and neurodegeneration diseases

The essential guide for extending your knowledge on the molecular mechanisms of neurodegenerative diseases - Guides

HTRF cellular phospho-protein assays

Physiologically relevant results fo fast flowing research - Flyers

Neurodegeneration and its main related diseases

Discover this infographic design on neurodegenerative diseases - Infographics

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