BRD4 Protein, Human (His-Flag)

The BRD4 protein is a chromatin reader that recognizes and binds acetylated histones, transmits epigenetic memory and regulates transcription during cell division. It associates with acetylated chromatin throughout the cell cycle, maintaining chromatin structure and contributing to epigenetic memory. BRD4 Protein, Human (His-Flag) is the recombinant human-derived BRD4 protein, expressed by E. coli , with N-10*His, N-Flag labeled tag.

BRD4 protein functions as a chromatin reader, specifically recognizing and binding acetylated histones, thereby playing a crucial role in transmitting epigenetic memory across cell divisions and regulating transcription. It remains associated with acetylated chromatin throughout the cell cycle, preserving the acetylated chromatin status and maintaining high-order chromatin structure, contributing to epigenetic memory for postmitotic G1 gene transcription. During interphase, BRD4 is pivotal in regulating the transcription of signal-inducible genes by associating with the P-TEFb complex and recruiting it to promoters. Additionally, it collaborates with JMJD6 in recruiting the P-TEFb complex to distal enhancers, known as anti-pause enhancers, displacing negative regulators and transforming P-TEFb into an active form that can phosphorylate the C-terminal domain of RNA polymerase II. BRD4 also acts as an atypical protein kinase, promoting phosphorylation of 'Ser-2' of the C-terminal domain of RNA polymerase II. Beyond histones, BRD4 recognizes and binds acetylated RELA, facilitating the recruitment of the P-TEFb complex and activating NF-kappa-B. Furthermore, it acts as a regulator of p53/TP53-mediated transcription, being recruited to specific target promoters upon phosphorylation by CK2. In the DNA damage response pathway, BRD4 acts as a chromatin insulator, inhibiting DNA damage response signaling by recruiting the condensin-2 complex to acetylated histones and limiting the spreading of histone H2AX/H2A.x phosphorylation. These diverse functions underscore the versatile role of BRD4 in epigenetic regulation, transcriptional control, and maintenance of chromatin structure.

1. Measured in a cell proliferation assay using M-NFS-60 cells. The ED₅₀ for this effect is ≤13.78 ng/mL, corresponding to a specific activity is ≥7.26×10⁴ units/mg.
2. Western blot analysis of protein Human SIRT5 (HY-P74545) (lane 2 (10ng) and lane 3 (20ng)) and Human BRD4 (HY-P7846) (lane 4 (10ng) and lane 5 (20ng)) using Flag Tag(HRP) (HY-P80111A) Mouse mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) in 5% non-fat milk in TBST at 4°C overnight.

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  Measured in a cell proliferation assay using M-NFS-60 cells. The ED₅₀ for this effect is 13.78 ng/mL, corresponding to a specific activity is 7.26×10⁴ units/mg.

Human

E. coli

N-10*His;N-Flag

O60885-1 (E49-E460)

23476  [NCBI]

Partial

ETSNPNKPKRQTNQLQYLLRVVLKTLWKHQFAWPFQQPVDAVKLNLPDYYKIIKTPMDMGTIKKRLENNYYWNAQECIQDFNTMFTNCYIYNKPGDDIVLMAEALEKLFLQKINELPTEETEIMIVQAKGRGRGRKETGTAKPGVSTVPNTTQASTPPQTQTPQPNPPPVQATPHPFPAVTPDLIVQTPVMTVVPPQPLQTPPPVPPQPQPPPAPAPQPVQSHPPIIAATPQPVKTKKGVKRKADTTTPTTIDPIHEPPSLPPEPKTTKLGQRRESSRPVKPPKKDVPDSQQHPAPEKSSKVSEQLKCCSGILKEMFAKKHAAYAWPFYKPVDVEALGLHDYCDIIKHPMDMSTIKSKLEAREYRDAQEFGADVRLMFSNCYKYNPPDHEVVAMARKLQDVFEMRFAKMPDE

Approximately 54-60 kDa

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  ≥ 90%, as determined by reducing SDS-PAGE.

Solution

1.Supplied as a 0.22 μm filtered solution of 50 mM HEPES, 200 mM NaCl, 10% glycerol, 1 mM DTT, pH 7.5.
2.Supplied as a 0.22 μm filtered solution of 50 mM HEPES, 300 mM NaCl, 1 mM DTT, 10% glycerol, pH 7.5.
3.Supplied as a 0.22 μm filtered solution of 50 mM HEPES, 300 mM NaCl, pH 7.4, 1 mM DTT, 10% glycerol.
4.Supplied as a 0.22 μm filtered solution of 50 mM HEPES, 300 mM NaCl, pH 7.4,1 mM DTT, 10% glycerol.
Please refer to the lot-specific COA for specific buffer information.

<1 EU/μg, determined by LAL method.

N/A.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.

• [1]. /biology-dictionary/sf9-cell.html  [Content Brief]