Revvity, 64BRD7TPEG, HTRF Human and Mouse Total BRD7 Detection Kit, 500 Assay Points

Overview
The Total BRD7 cellular assay monitors BRD7 protein levels.
Bromodomain containing 7 (BRD7), also known as NAG4, BP75 or CELTIX1, is a member of the bromodomain-containing protein family IV, which also includes BRD9 protein. BRD7 is mainly localized in the nucleus and regulates chromatin remodeling. BRD7 is a component of a large polybromo-BAF (PBAF) complex, including ARID2, PBRM1, and BAF45d, which interacts with transcriptional regulators to modulate gene expression.
More importantly, recent studies have shown that BRD7 is downregulated in certain cancers such as epithelial ovarian carcinoma, breast cancer, nasopharyngeal cancer, and colorectal carcinoma. Unlike prooncogenic BRD9 roles, BRD7 may function as a tumor suppressor by inhibiting the Phosphoinositide 3-kinase (PI3K) pathway involved in various deregulated oncogenic processes, such as apoptosis, protein synthesis, proliferation, and angiogenesis.
Since BRD7 and BRD9 proteins share a 72% residue similarity, compounds are likely to target both the tumor suppressor BRD7 and the prooncogene BRD9. 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-BRD7 assay principle
The HTRF Total-BRD7 assay quantifies the expression level of BRD7 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-BRD7 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 BRD7 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-BRD7 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-BRD7 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total-BRD7 one-plate assay protocol
Detection of Total-BRD7 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 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-500 assay points