| 货号 | 13440S |
| 反应种属 | Human |
| 来源宿主 | Rabbit |
| 应用 | W/IP/ChIP |
| 目标/特异性 | BRD4 (E2A7X) Rabbit 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. |
| 使用方法 | WB(1:1000) IP (1:50) ChIP (1:50) |
| 供应商 | CST |
| 背景 | 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). |
| 存放说明 | -20C |
| 计算分子量 | 200 |
Western blot analysis of extracts from RL-7, 293T, and Jurkat cells using BRD4 (E2A7X) Rabbit mAb.Western blot方法检测RL-7, 293T,以及Jurkat细胞系的提取物，使用的抗体为BRD4 (E2A7X) Rabbit mAb。 | |
Immunoprecipitation of BRD4 from RL-7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or BRD4 (E2A7X) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using BRD4 (E2A7X) Rabbit mAb.从RL-7细胞提取物中免疫沉淀BRD4，使用的抗体为Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) 或BRD4 (E2A7X) Rabbit mAb (lane 3)。Lane 1为10% input。 使用BRD4 (E2A7X) Rabbit mAb进行Western blot检测。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106MV-4-11 cells and either 10 µl of BRD4 (E2A7X) Rabbit mAb or 2 µl of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Bcl-2 Promoter Primers #12924, human c-Myc intron 1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对4 x 106 MV-4-11细胞中交联过的染色质进行染色质免疫沉淀，所用抗体为10 µl BRD4 (E2A7X) Rabbit mAb 或2 µl Normal Rabbit IgG #2729，富集到的DNA经过实时PCR定量，所使用产品为SimpleChIP® Human Bcl-2 Promoter Primers #12924, human c-Myc intron 1 primers以及SimpleChIP® Human α Satellite Repeat Primers #4486。每个样品中免疫沉淀得到的DNA量由与input chromatin( 相当于1)的相对信号值来表示。 |
