Revision 8
#5848
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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
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
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
- Florence, B. and Faller, D.V. (2001) Front Biosci 6, D1008-18.
- Denis, G.V. et al. (2000) Cell Growth Differ 11, 417-24.
- Crowley, T.E. et al. (2002) Mol Endocrinol 16, 1727-37.
- Denis, G.V. et al. (2006) J Proteome Res 5, 502-11.
- Gyuris, A. et al. (2009) Biochim Biophys Acta 1789, 413-21.
- Kanno, T. et al. (2004) Mol Cell 13, 33-43.
- Shang, E. et al. (2004) Gene Expr Patterns 4, 513-9.
- Trousdale, R.K. and Wolgemuth, D.J. (2004) Mol Reprod Dev 68, 261-8.
- Pal, D.K. et al. (2003) Am J Hum Genet 73, 261-70.
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 ChIP: Chromatin IP C&R: CUT&RUN
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.
Limited Uses
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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 8
#5848
Brd2 (D89B4) Rabbit mAb
Western blot analysis of extracts from MOLT-4 and NCCIT cells using 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.
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.
Revision 8
#5848
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 chromosome 6 (upper), including GTF3C6 (lower), a known target gene of BRD2 (see additional figure containing ChIP-qPCR data).
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.
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.
Revision 8
#5848
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 figures show binding across HOXD genes (upper) and H2BC genes (lower).
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.