Revvity, 64BRD9TPEH, HTRF Human and Mouse Total BRD9 Detection Kit, 10,000 Assay Points

Overview
BRD9 (BRD9 Bromodomain-containing protein 9) is a member of the bromodomain-containing protein family IV which also includes BRD7. BRD9 is a SWI/SNF (GBAF or ncBAF) subunit and binds acetylated lysine residues of histones, mediating chromatin remodeling and transcriptional regulators. BRD9 is involved in pluripotency regulation and cell development.
BRD9 overexpression has been associated with neurodevelopmental disorders and cancers such as Acute Myeloid Leukemia. Targeting BRD9 degradation via PROTAC molecules has recently emerged as a novel and promising therapeutic approach. However, BRD9 and BRD7 proteins share 72% residue similarity, thus compounds are likely to target both the pro-oncogene BRD9 and the tumor suppressor BRD7. Therefore, the selectivity of compounds targeting either BRD9 or BRD7 must be carefully investigated. To do this, the HTRF total BRD7 kit is a valuable companion assay for the HTRF total BRD9 kit, enabling reliable compound profiling.
How it works
Total-BRD9 assay principle
The HTRF Total-BRD9 assay quantifies the expression level of BRD9 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-BRD4 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 the presence of BRD9 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-BRD9 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-BRD9 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total-BRD9 one-plate assay protocol
Detection of Total-BRD9 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 Mouse
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-10,000 assay points