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63759
BRD4 (E4X7E) Mouse mAb
Primary Antibodies
Monoclonal Antibody

BRD4 (E4X7E) Mouse mAb #63759

Citations (1)
Western blot analysis of extracts from various cell lines using BRD4 (E4X7E) Mouse mAb.
Simple Western analysis of lysates (1.0 mg/mL) from Jurkat cells using BRD4 (E4X7E) Mouse mAb #63759. The virtual lane view (left) shows the target bands (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess ​​​​​​​Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
Immunoprecipitation of BRD4 protein from 293T cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is BRD4 (E4X7E) Mouse mAb. Western blot analysis was performed using BRD4 (E4X7E) Mouse mAb.
Confocal immunofluorescent analysis of active transcription bodies within a HeLa cell labeled with BRD4 (E4X7E) Mouse mAb (green, left) and colabeled with DAPI #4083 (blue, right).
Immunofluorescent analysis of HCT 116 cells, either mock transfected (green, high-expressing) or transfected with siRNA directed against BRD4 (red, low-expressing), using BRD4 (E4X7E) Mouse mAb. Various concentrations of antibody (x-axis) were tested and fluorescence intensities were measured to determine specificity and optimal working concentration.
Confocal immunofluorescent analysis of HCT 116 cells, either mock transfected (left, high-expressing) or transfected with siRNA directed against BRD4 (right, low-expressing), using BRD4 (E4X7E) Mouse mAb (green) and DAPI #4083 (blue).

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:6400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

BRD4 (E4X7E) Mouse mAb recognizes endogenous levels of total BRD4 protein. This antibody specifically recognizes the BRD4 long isoform (UniProt #O60885-1) and does not recognize other BRD4 isoforms.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu1129 of human BRD4 protein.

Background

Bromodomain-containing protein 4 (BRD4) is a member of the bromodomains and extra terminal (BET) family of proteins, which also includes BRD2, BRD3, and BRDT (1-3). BET family proteins contain two tandem bromodomains and an extra terminal (ET) domain, and bind acetyl lysine residues (3). BRD4 is a chromatin-binding protein with a preference for Lys14 on histone H3 as well as Lys5 and Lys12 on histone H4 (4). BRD4 chromatin binding occurs throughout the cell cycle, including condensed mitotic chromosomes, when the majority of genes are silenced (5). BRD4 association with chromatin during mitosis is thought to be an important part of the bookmarking mechanism to accelerate re-activation of the silenced genes upon exit from mitosis (2,6). BRD4 has been shown to facilitate transcription by recruiting the positive transcription elongation factor b (pTEFb) complex that phosphorylates Ser2 of the heptapeptide repeat of the carboxy-terminal domain of RNA polymerase II, promoting transcription elongation (3,7,8). In addition, BRD4 has been found to be part of the super elongation complex and the polymerase associated factor complex (PAFc) in MLL-fusion derived leukemia cell lines, demonstrating a role for BRD4 in the regulation of transcription elongation (9). Research studies have shown that BRD4 (and BET family proteins) may be promising therapeutic targets for various Myc-driven cancers, such as Burkitt’s lymphoma and certain acute myeloid leukemias (1,10,11). Investigators have found molecular inhibition of BET proteins to be effective in inducing apoptosis in various MLL-fusion driven leukemic cell lines by competing BRD3 and BRD4 from chromatin, leading to reduced expression of Bcl-2, Myc, and CDK6 (9). BET inhibition has also been shown to have antitumor activities against nuclear protein in testis (NUT) midline carcinoma cell lines and xenografts in mice where BRD4 is found to be a frequent translocation partner of the NUT protein (12). In addition, BRD4 regulates the expression of some inflammatory genes, and inhibition of BRD4 (and BET family proteins) chromatin binding causes reduced expression of a subset of inflammatory genes in macrophages, leading to protection against endotoxic shock and sepsis (13).

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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To Purchase # 63759
Cat. # Size Qty. Price
63759S
100 µl