Revvity, 64NTBPEH, HTRF Human and Mouse Total TBK1 Detection Kit, 10,000 Assay Points

Overview
The Total TBK1 cellular assay is designed to monitor the expression level of TBK1 proteins, both phosphorylated and unphosphorylated. It is compatible with our phospho-TBK1 kit and enables the analysis of phosphorylated and total proteins from a single sample for a better readout of T-cell activation. Following pathogen infection, TBK1 can be activated by bacterial LPS and pathogen nucleic acids which trigger the activation of TLR3/4 and cGAS-STING signaling axis.
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
Total TBK1 assay principle
The total TBK1 assay quantifies the expression level of TBK1 in a cell lysate. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The total TBK1 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 TBK1 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 proteins expression under a no-wash assay format.

 

Total TBK1 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 adding Total TBK1 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total TBK1 1-plate assay protocol
Detection of total TBK1 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 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 Mouse
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-10,000 assay points