Revision 8

#5848

Store at -20C

Brd2 (D89B4) Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Applications:
W, W-S, ChIP, ChIP-seq, C&R

Reactivity:
H M

Sensitivity:
Endogenous

MW (kDa):
110

Source/Isotype:
Rabbit IgG

UniProt ID:
#P25440

Entrez-Gene Id:
6046

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 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

Brd2 (D89B4) Rabbit mAb recognizes endogenous levels of total Brd2 protein. This antibody may cross-react with Brd3 protein.

Species predicted to react based on 100% sequence homology

Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala310 of human Brd2 protein.

Background

Brd2 is a highly conserved member of the BET subfamily of bromodomain proteins that contain two tandem N-terminal bromodomains and a single C-terminal extra-terminal (ET) domain (1). In addition to its involvement in guiding the expression of cell cycle genes through its binding to multiple E2Fs (2), Brd2 has been shown to be associated with several regulators of transcription, including TFIID and Swi/Snf complexes (3,4). First identified as a nuclear serine/threonine kinase (2), Brd2, like other bromodomain proteins, is thought to function in mammalian development by regulating chromatin structure and transcription (5). Brd2 has been shown to bind to histone H4 via acetylated Lys12, a substrate of several histone acetyltransferase transcriptional coactivators (6). In mouse, Brd2 has the highest levels of expression during embryogenesis and in the adult testis, ovaries, and brain (3,7,8). Brd2-deficient mouse embryos exhibit delayed development and eventual death due to neural tube closure defects (5). Mutations in the promoter of the Brd2 gene have been associated with increased susceptibility to juvenile myoclonic epilepsy (JME) (9).

Background References

Species Reactivity

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

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

Cross-Reactivity Key

H: Human M: Mouse

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 8

#5848

Brd2 (D89B4) Rabbit mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from MOLT-4 and NCCIT cells using Brd2 (D89B4) Rabbit mAb.
Western Blotting Image 1: Brd2 (D89B4) Rabbit mAb
Simple Western™ analysis of lysates (0.1 mg/mL) from Molt-4 cells using Brd2 (D89B4) Rabbit mAb #5848. The virtual lane view (left) shows the target band (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 12-230 kDa separation module.
Western Blotting Image 1: Brd2 (D89B4) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and Brd2 (D89B4) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across GTF3C6, a known target gene of BRD2 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin Immunoprecipitation Image 1: Brd2 (D89B4) 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 8

#5848

Brd2 (D89B4) Rabbit mAb

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and Brd2 (D89B4) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 6 (upper), including GTF3C6 (lower), a known target gene of BRD2 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: Brd2 (D89B4) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either Brd2 (D89B4) 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 GTF3C6 exon 1 primers, SimpleChIP® Human SF3B3 Exon 1 Primers #62858, and SimpleChIP® Human MyoD1 Exon 1 Primers #4490. 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: Brd2 (D89B4) Rabbit mAb
CUT&RUN was performed with NCCIT cells and Brd2 (D89B4) Rabbit mAb, using CUT&RUN Assay Kit #86652. The DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HOXD genes.
CUT & RUN Image 1: Brd2 (D89B4) 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 8

#5848

Brd2 (D89B4) Rabbit mAb

Cell Signaling Technology Logo
CUT&RUN was performed with NCCIT cells and Brd2 (D89B4) Rabbit mAb, using CUT&RUN Assay Kit #86652. The DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across HOXD genes (upper) and H2BC genes (lower).
CUT & RUN Image 2: Brd2 (D89B4) Rabbit mAb
CUT&RUN was performed with NCCIT cells and either Brd2 (D89B4) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human LPAR3 promoter primers and human POTEKP promoter primers. 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.
CUT & RUN Image 3: Brd2 (D89B4) 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.