IMP-6617

BRD4

The peptide corresponding to amino acids 333-460 that contains the bromodomain sequences of BRD4 (accession number NP_490597.1) was expressed in E. coli and contains an N-terminal His-Tag and C-terminal FLAG-Tag with an observed molecular weight of 20.9 kDa. The recombinant protein is >87% pure by SDS-PAGE.

BRD4 (Bromodomain-containing protein 4) belongs to the BET subclass of proteins, which are characterized by two N-terminal bromodomains and one ET (Extra Terminal) domain. BRDs associate with chromatin through their bromodomains that recognize acetylated histone lysine residues. Bromodomains function as 'readers' of these epigenetic histone marks and regulate chromatin structure and gene expression by linking associated proteins to the acetylated nucleosomal targets. The ET domain functions as a protein binding motif and exerts atypical serine-kinase activity. The BET family consists of at least four members in mouse and human, BRD2 (also referred to as FSRG1, RING3), BRD3 (FSRG2, ORFX), BRD4 (FSRG4, MCAP/HUNK1), and BRDT (FSRG3, BRD6). BRD proteins are related to the female Sterile Homeotic protein gene in Drosophila, a gene required maternally for proper expression of other homeotic genes, such as Ubx, which is involved in pattern formation. BRD4 has been identified recently as a therapeutic target in many cancers, including acute myeloid leukemia, multiple myeloma, Burkitt's lymphoma, NUT midline carcinoma, colon cancer, and breast cancer. BRD4 regulates the transcription of oncogenes, HIV, and human papilloma virus (HPV). It has been shown to bind and phosphorylate RNA pol II, which implicates its involvement in the regulation of eukaryotic transcription.

E. coli

Human

His/FLAG

Recombinant BRD4 (333-460) protein expressed in E. coli and supplied in 25 mM HEPES pH 8, 500 mM NaCl and 20% glycerol.

20.9 kDa

333-460

>87%

Where possible, Active Motif has developed functional or activity assays for recombinant proteins. Additional characterization such as enzyme kinetic activity assays, inhibitor screening or other biological activity assays may not have been performed for every product.

Recombinant proteins in solution are temperature sensitive and must be stored at -80 centigrade to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Recombinant BRD4 (333-460) protein gel. BRD4 (333-460) protein was run on a 12.5%SDS-PAGE gel and stained with Coomassie blue. MW: 20.9 kDa Purity: >92%

HTRF assay for BRD4 (333-460) activity 3 μM histone peptide H4K5/8/12/16(ac4) was incubated with BRD4 (333-460) in a reaction buffer including 50 mM HEPES-NaOH pH 7.0, and 0.1% BSA for 1 hour at room temperature. Anti-FLAG antibody was used to detect reaction products.