Revvity, 64BRD3TPEH, HTRF Human Total BRD3 Detection Kit, 10,000 Assay Points

Overview
BRD3 belongs to the bromodomain (BRD) Family II (BET). Like the other BRD family proteins, it binds to acetylated lysine and acts as a reader of the lysine acetylation state of proteins such as histones. It is a transcriptional regulator involved in many cell processes (embryogenesis, cycle regulation, development) through the E2F-RB pathway. BRD3 interacts with the GATA transcription factor for its chromatin occupancy and the regulation of several oncogenes, such as c-MYC.
Thus, inhibition of BRD3 function has become part of the strategy for cancer treatments. More recently, targeting BET family protein degradation, including BRD3 via PROTAC molecules, has emerged as a novel and promising therapeutic approach.
How it works
Total-BRD3 assay principle
The HTRF Total-BRD3 assay quantifies the expression level of BRD3 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-BRD3 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 BRD3 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-BRD3 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of Total-BRD3 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total-BRD3 one-plate assay protocol
Detection of Total-BRD3 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
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4
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-Oncology & Inflammation
Unit Size-10,000 assay points