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Recombinant Proteins & Enzymes

Recombinant BRD4 (44-460) protein

Catalog No: 31594 Format: 100 µg $395 Buy Now
Catalog No: 31994 Format: 1 mg $2,600 Buy Now
Expressed In: E. coli Protein Species: Human

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for Recombinant BRD4 (44-460) proteinContents

Recombinant BRD4 (44-460) protein is supplied in 25 mM Tris pH 8.0, 300 mM NaCl, 10% glycerol, 0.5 mM TCEP. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant BRD4 (44-460) proteinBackground

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.

for Recombinant BRD4 (44-460) proteinApplication Notes

This protein is suitable for use in binding assays, inhibitor screening, and selectivity profiling.

for Recombinant BRD4 (44-460) proteinProtein Details

The protein corresponding to amino acids 44 - 460 that contains two Bromo domains of BRD4 (accession number NP_490597.1) was expressed in E. coli cells and contains an N-terminal His tag and a C-terminal FLAG tag. The molecular weight of BRD4 (44-460) is 51.6 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.

 
Recombinant BRD4 (44-460) protein gel

Recombinant BRD4 (44-460) protein gel
10% SDS-PAGE gel, stained with Coomassie blue.
MW: 51.6 kDa
Purity: > 85%

HTRF assay for BRD4 (44-460) activity

HTRF assay for BRD4 (44-460) activity
1 µM H4K5/8/12/16(4ac) peptide was incubated with different concentrations of BRD4 (44-460) protein in 10 µl reaction system containing 50 mM HEPES-NaOH pH 7.5, 0.1% BSA for 1 hour, then 10 µl Anti-FLAG antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) were added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. 

HTRF assay for BRD4 (44-460) activity

HTRF assay for BRD4 (44-460) activity
1 µM H4K5/8/12/16(4ac) peptide was incubated with different concentrations of BRD4 (44-460) protein in 10 µl reaction system containing 50 mM HEPES-NaOH pH 7.5, 0.1% BSA for 1 hour, then 10 µl Anti-FLAG antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) were added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. 

for Recombinant BRD4 (44-460) proteinStorage

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.

for Recombinant BRD4 (44-460) proteinGuarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.

 

Technical Data Sheet

Recombinant BRD4 (44-460) protein
 

Data Thumbnails

Recombinant BRD4 (44-460) protein gel

Recombinant BRD4 (44-460) protein gel
(Click to enlarge and view details)

HTRF assay for BRD4 (44-460) activity

HTRF assay for BRD4 (44-460) activity
(Click to enlarge and view details)

HTRF assay for BRD4 (44-460) activity

HTRF assay for BRD4 (44-460) activity
(Click to enlarge and view details)