Recombinant BRD4 (44-168) protein

Aliases: CAP; MCAP; HUNK1; HUNKI
Catalog No: 31380 Format: 100 µg $405 Buy
Catalog No: 31880 Format: 1 mg $2,600 Buy
Expressed In: E. coli Protein Species: Human

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Contents

Recombinant BRD4 (44-168) protein was expressed in E. coli and is supplied in 25 mM Tris pH 8.0, 300 mM NaCl and 5% glycerol. Please refer to product insert upon arrival for lot-specific concentration.

Background

Bromodomain-containing protein 4 (BRD4) 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.

Application Notes

Recombinant BRD4 (44-168) is suitable for use in binding assays, inhibitor screening, and selectivity profiling.

Protein Details

The peptide corresponding to amino acids 44 - 168 that contains the bromodomain sequences of BRD4 (accession number NM_058243.2) was expressed in E. coli and contains an N-terminal His-Tag and C-terminal FLAG-Tag with an observed molecular weight of 20 kDa. It shows binding specificity for acetylated H3K9, H3K9/K14, H4K5, H4K8, H4K12, H4K5/K8, H4K5/K12, H4K8/K12, H4K12/K16, H4K12/K16/K20 and H4K5/K8/K12/K16, as well as acetylated RelA-K310. The recombinant protein is >90% pure by SDS-PAGE.

References


This product was used in the following publications:
Bioorg. Med. Chem. (2017). 25(8):2482-2490. PMID: 28314513.

 

Recombinant BRD4 (44-168) protein gel.
BRD4 (44-168) protein was run on an SDS-PAGE gel and stained with Coomassie Blue.

Recombinant BRD4 (44-168) HTRF activity assay.
3.3 uM histone peptide H4K5/8/12/16(4Ac) was incubated with BRD4 (44-168) protein in reaction buffer including 50mM HEPES-NaOH pH 7.0, 0.1% BSA for 1 hour at room temperature. Anti-Flag antibody was used to detect reaction products

Storage

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

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.