Recombinant Human BRD4 Protein (333-460), GST-tagged
Product Description
Cat
IMP-6614
Official Symbol
BRD4
Product Overview
Recombinant BRD4 (333-460), GST-Tag protein that includes amino acids 333-460 (the second Bromodomain) of human BRD4 protein (accession number NP_490597.1) was expressed in E.coli and contains an N-terminal GST Tag with a molecular weight of 41.1 kDa.
Description
(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.
Expression System
E. coli
Species
Human
Tag
GST
Form
Recombinant BRD4 (333-460), GST-Tag protein is supplied in 25 mM Tris-HCl pH 8.0, 300 mM NaCl, 10% glycerol.
Molecular Mass
41.1 kDa
Protein length
333-460
Purity
>91%
Applications
Binding assays, inhibitor screening, and selectivity profiling
Storage
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.
SDS-PAGE
SDS-PAGE

Recombinant BRD4 (333-460), GST-Tag protein gel 10% SDS-PAGE Coomassie staining MW: 41.1 kDa Purity: >91%

Bioactivity-ELISA 1
Activity 1

HTRF assay for BRD4 (333-460), GST-Tag activity 1 μM H4K5/8/12/16(ac4) peptide was incubated with different concentrations of BRD4 (333-460), GST-Tag protein in a 10 μL binding system containing 50 mM HEPES-NaOH pH 7.4, 0.1% BSA for 1 hour, then 10 μL GST antibody and SA-XL665 mixture (each 1:100 dilution in Binding Buffer) were added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

Bioactivity-ELISA 2
Activity 2

HTRF assay for BRD4 (333-460), GST-Tag activity 1 μM H4K5/8/12/16(ac4) peptide was incubated with different concentrations of BRD4 (333-460), GST-Tag protein in a 10 μL binding system containing 50 mM HEPES-NaOH pH 7.4, 0.1% BSA for 1 hour, then 10 μL GST antibody and SA-XL665 mixture (each 1:100 dilution in Binding Buffer) were added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

Data Sheet MSDS
For Research Use Only. Not intended for any clinical use. No products from Creative BioMart may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative BioMart.

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