Revvity, 64TDPS4PEH, HTRF Human and Mouse Phospho-TDP-43 (Ser409) Detection Kit, 10,000 Assay Points

Overview
TAR DNA binding protein 43 (TDP-43) is a nucleic acid binding protein involved in RNA-related metabolism. Aggregated TDP-43 has been identified as a hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia (FTLD), and more widely in several neurodegenerative diseases: TDP-43 proteinopathies. In pathological conditions (mutation or dysregulation), TDP-43 forms insoluble inclusion bodies in the cytoplasm of neurons in the brain and spinal cord. The TDP-43 Phospho-Ser409/410 assay detects human TDP-43 phosphorylated in cell lysates.
How it works
Phospho-TDP-43 (Ser409/410) assay principle
The Phospho-TDP-43 (Ser409/410) assay measures TDP-43 when phosphorylated at Ser409/410. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-TDP-43 (Ser409/410) assay uses 2 antibodies, one labeled with a donor fluorophore and 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 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-TDP-43 (Ser409/410) two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates to a 384-well low volume detection plate before adding Phospho-TDP-43 (Ser409/410) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

Phospho-TDP-43 (Ser409/410) one-plate assay protocol
Detection of Phosphorylated TDP-43 (Ser409/410) 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 4
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
Unit Size-10,000 assay points