Revvity, 62TDP43PEG, HTRF TDP-43 Aggregation Kit, 500 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 deregulation), TDP-43 forms insoluble inclusion bodies in the cytoplasm of neurons in the brain and spinal cord. The TDP-43 aggregation assay detects human and mouse TDP-43 aggregates rapidly in cellular models.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
TDP-43 aggregation assay principle
TDP-43 aggregates are measured using a comparison between two HTRF specific signals: the control condition, where soluble TDP-43 only is detected, and the disaggregated condition, where total disaggregated and soluble TDP-43 are assessed. The increase in the disaggregated HTRF signal reflects the level of TDP-43 aggregation.

 

TDP-43 disaggregation step
Samples containing TDP-43 aggregates require a disaggregation step in order to calculate the level of aggregation of TDP-43 between 2 conditions prior and after sample treatment (sample disaggregation can follow the protocol described on the right). The reagents needed for this step are provided in the kit for maximum convenience. Samples are dispensed directly into the detection (white, low-volume) plate for detection by HTRF® reagents.

 

TDP-43 aggregation detection
The same control condition sample treated or after the disaggregation step is dispensed directly into the assay plate for detection by HTRF® reagents (384-well low-volume white plate or Revvity low-volume 96-well plate in 20 µl). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up to a 1536 well-format by simply resizing each addition volume proportionally.

 

TDP-43 aggregation assay data analysis
The aggregated ratio of each sample reflects the presence of TDP-43 aggregates in a sample.
In the case of absence of insoluble aggregates, the aggregated ratio is around 1 or below. Application-Protein Quantification
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Biomarkers
Technology-TR-FRET
Therapeutic Area-Neuroscience
Unit Size-500 assay points