Skip to main content
Menu
US

HTRF Human Phospho-Tau (Ser202/Thr205) Detection Kit, 500 Assay Points

This HTRF kit enables the cell-based detection of phosphorylated TAU at Ser202/Thr205, as a marker of neurodegenerative diseases.

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 & disposal requirements under European REACH regulations (EC 1907/2006).

Product Variants
Part number: 64TS2PEG
Unit Size: 500 Assay Points
List price: USD 2,147.00
Your price: Login to view
USD 2,147.00
USD 2,147.00 /each
Part number: 64TS2PEH
Unit Size: 10,000 Assay Points
List price: USD 12,490.00
Your price: Login to view
USD 0.00
USD 12,490.00 /each

Overview

The HTRF Phospho-TAU (Ser202/Thr205) cell-based assay kit is ideal for quantifying endogenous phospho-TAU phosphorylated on Serine 202/Threonine 205.

TAU exists in different states in both Alzheimer’s (AD) and Parkinson’s (PD) diseases. Tau hyperphosphorylation is also a marker for multiple neurodegenerative diseases and CNS disorders.

Specifications

Assay Points
500
Assay Technology
HTRF
Brand
HTRF
Product Group
Kit
Quantity
1
Shipping Conditions
Shipped in Dry Ice
Therapeutic Area
Neuroscience
Unit Size
500 Assay Points

Video gallery

Citations

How it works

Phospho-TAU (Ser202/Thr205) assay principle

The Phospho-TAU (Ser202/Thr205) assay measures TAU when phosphorylated at Ser202/Thr205. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer.

The Phospho-TAU (Ser202/Thr205) assay uses 2 labeled antibodies: one with a donor fluorophore, the other with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, and the second for its ability to recognize the protein independently of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both 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 assay principle
Phospho-TAU (Ser202/Thr205) 2-plate assay protocol

The 2 plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates to a 384-well low volume detection plate before the additio,n of Phospho-TAU (Ser202/Thr205) HTRF detection reagents.

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

Assay principle
Phospho-TAU (Ser202/Thr205) 1-plate assay protocol

Detection of Phosphorylated TAU (Ser202/Thr205) 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 protocol

 

Assay validation

Inhibition of GSK3b impairs phosphorylation of TAU on S202/T205 residues

50,000 human SH-SY5Y cells were plated in 96-well plates and incubated for 72h at 37 °C - 5% CO2.

After incubation with increasing concentrations of GSK3 inhibitors (CHIR99021 and BIO) for 3 hours, medium was removed and cells were lysed with 50µl of lysis buffer for 30min at RT under gentle shaking. 16 µL of lysate were transferred into a 384-well sv white microplate, and 4 µL of the HTRF phospho-TAU (Ser202/Thr205) detection reagents were added.

The HTRF signal was recorded after an overnight incubation.

Assay validation tau phospho
Phosphorylation of TAU on Ser202/Thr205 is abrogated by alkaline Phosphatase treatment

50,000 human SH-SY5Y cells were plated in 96-well plates and incubated for 72h at 37 °C - 5% CO2. Medium was removed and cells were lysed with 50µl of lysis buffer for 30min at RT under gentle shaking.

Alcaline phosphatase (25U) was added with the appropriate buffer and removed 1 hour later. 16 µL of lysate were transferred into a 384-well sv white microplate, and 4 µL of the HTRF phospho-TAU (Ser202/Thr205) detection reagents were added. The HTRF signal was recorded after an overnight incubation.

Assay validation tau phospho
BIO inhibitor leads to inhibition of TAU phosphorylation, whereas a-tubulin remains constant

50,000 human SH-SY5Y cells were plated in 96-well plates and incubated for 72h at 37 °C - 5% CO2. The SH-SY5Y cells were then treated with the GSK3 inhibitor, BIO, for 3 hours. After cell culture medium removal, cells were lysed with 50µl of lysis buffer for 30min at RT under gentle shaking. 16 µL of lysate were transferred into a 384-well sv white microplate, and 4 µL of the HTRF phospho-TAU (Ser202/Thr205) and α-tubulin detection reagents were added.

The HTRF signal was recorded after an overnight incubation.

Phospho s202 t205
HTRF vs Western blot comparison

Human SH-SY5Y cells were grown in a T175 flask at 37 °C, 5% CO2 until 80% confluency. Cell culture medium was discarded, and cells were lysed with 3 mL of supplemented lysis buffer for 30 min at room temperature. Soluble fractions were then collected after a 10 min centrifugation. Serial dilutions of the cell lysate were performed in the supplemented lysis buffer, and then 16 µL of each dilution were dispensed and analyzed side-by-side by Western Blot and by HTRF.

Using the HTRF phospho-TAU (Ser202/Thr205) cellular assay, just 2,500 cells were sufficient for minimum signal detection, while 5,000 cells were needed for a Western Blot signal. The HTRF cellular assays are at least 2-fold more sensitive than the Western Blot.

Assay validation tau phospho

 

Simplified pathway

Alzheimer's disease pathway and TAU role

TAU has a prominent role in the pathogenesis of Alzheimer's disease. It becomes hyperphosphorylated and aggregates, forming filaments, which can further condense into neurofibrillary tangles. TAU aggregates may propagate pathology by spreading from cell to cell in a prion-like manner. Drugs modulating TAU hyperphosphorylation and reducing TAU aggregation are viable therapeutic approaches.

The physiological role of TAU protein is to promote assembly and stability of microtubules. Six isoforms of TAU have been described, ranging from 352 to 441 residues coming from exons 2, 3 and 10, that are alternatively spliced. The longest isoform of TAU (TAU-441) contains 85 putative phosphorylation sites, half of which have been confirmed experimentally.

Phospho pathway tau phospho

 

Resources

1-2 of 2 Resources
Brochure Icon
Brochure
HTRF assays and reagents catalog

Discover the versatility and precision of Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our HTRF portfolio offers a...

Guide Icon
Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

SDS, COAs, Manuals and more

Are you looking for technical documents related to the product? We have categorized them in dedicated sections below. Explore now.

Safety data sheet
+ Show more
Certificate of analysis
+ Show more
IFUs, Manuals & more
Scroll Icon