Revision 3

#94032

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, ChIP, ChIP-seq

Reactivity:
H Mk

Sensitivity:
Endogenous

MW (kDa):
100

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q15059

Entrez-Gene Id:
8019

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Chromatin IP 1:50
Chromatin IP-seq 1:50

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

BRD3 (E3D5N) Rabbit mAb recognizes endogenous levels of total BRD3 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala10 of human BRD3 protein.

Background

Bromodomain-containing protein 3 (BRD3), also known as ORFX, is a member of the bromodomains and extra terminal (BET) family of proteins, which also includes BRD2, BRD4, and BRDT (1-3). BET family proteins contain two tandem bromodomains and an extra terminal (ET) domain, and bind acetyl lysine residues (3). BRD3 is capable of binding acetylated histone H3 Lys14 as well as acetylated histone H4 Lys5 and Lys12 to promote transcription (4). BRD3 plays a role in erythroid development by binding to GATA1 to facilitate its binding to target genes (5). Similar to BRD4, the BRD3 gene can be fused to NUT in NUT midline carcinomas (6). 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 BCL2, Myc, and CDK6 (7).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

W: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP

Cross-Reactivity Key

H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 , (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

CST Logo
Western blot analysis of extracts from various cell lines using BRD3 (E3D5N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Low expression of BRD3 protein in NCI-H929 cells is consistent with the predicted expression pattern.
Western Blotting Image 1: BRD3 (E3D5N) Rabbit mAb
Western blot analysis of extracts from 293T cells, transfected with control siRNA (-) or human BRD3 siRNA (hBRD3 siRNA; +), using BRD3 (E3D5N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: BRD3 (E3D5N) Rabbit mAb
Immunoprecipitation of BRD3 protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is BRD3 (E3D5N) Rabbit mAb. Western blot analysis was performed using BRD3 (E3D5N) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: BRD3 (E3D5N) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

CST Logo

Chromatin immunoprecipitations were performed with cross-linked chromatin from MV-4-11 cells and BRD3 (E3D5N) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across Myc, a known target gene of BRD3 (see additional figure containing ChIP-qPCR data).

Chromatin Immunoprecipitation Image 1: BRD3 (E3D5N) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from MV-4-11 cells and BRD3 (E3D5N) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across MYC (upper) and BCL2 (lower), known target genes of BRD3 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: BRD3 (E3D5N) Rabbit mAb

Chromatin immunoprecipitations were performed with cross-linked chromatin from MV-4-11 cells and either BRD3 (E3D5N) Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR, using human c-Myc promoter primers, human BCL2 promoter 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.

Chromatin Immunoprecipitation Image 3: BRD3 (E3D5N) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.