Revvity, 64NTAUPEG, HTRF Human Total Tau Detection Kit, 500 Assay Points

Overview
The Total Tau cell-based assay is designed to monitor the expression level of Tau proteins, both phosphorylated and unphosphorylated. It uses the same buffers as our Phospho-Tau kits, and enables the analysis of phosphorylated and total proteins from a single sample.
TAU (Tubulin Associated Units), also called MAPT (Microtubule Associated Protein Tau), is a phosphoprotein largely expressed in neurons of the Central Nervous System and involved in microtubule assembly and stability.
Tau contains around of 85 potential phosphosites that can be largely phosphorylated by many kinases such as CDK5, MAP kinase, or GSK3β, the main kinase of Tau. Tau hyperphosphorylation induces a microtubule disruption, Tau aggregation into paired helical filaments or neurofibrillary tangles, and cell death.
Tau hyperphosphorylation and aggregation are involved in numerous neurodegenerative diseases such as Tauopathy, and Parkinson's and Alzheimer's Diseases. These two phenomena associated with amyloid-β aggregation are major hallmarks of Alzheimer's Disease.
How it works
Total Tau assay principle
The Total Tau assay quantifies the expression level of Tau in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer.
The Total Tau 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 Tau 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 tau 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 Tau HTRF detection reagents.
This protocol enables the cells' viability and confluence to be monitored.

 

Total Tau 1-plate assay protocol
Detection of total Tau 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. 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-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Neuroscience
Unit Size-500 assay points