Rabbit Polyclonal BRD4 antibody. Suitable for IHC-P, IP, WB and reacts with Mouse, Human samples. Immunogen corresponding to Synthetic Peptide within Human BRD4 aa 1300 to C-terminus.
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Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7 - 8
Preservative: 0.09% Sodium azide
Constituents: Tris citrate/phosphate- Form
- Liquid
- Clonality
- Polyclonal
Immunogen
- Synthetic Peptide within Human BRD4 aa 1300 to C-terminus. The exact immunogen used to generate this antibody is proprietary information. Database link O60885
Reactivity data
Select an application| IHC-P | IP | WB | |
|---|---|---|---|
| Human | Tested | Tested | Tested |
| Mouse | Tested | Expected | Tested |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1/1000.00000 - 1/5000.00000 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/1000.00000 - 1/5000.00000 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 2.00000-5.00000 µg/mg of lysate | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1/2000.00000 - 1/10000.00000 | Notes - |
Species Human | Dilution info 1/2000.00000 - 1/10000.00000 | Notes - |
Associated Products
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Target data
Function
Chromatin reader protein that recognizes and binds acetylated histones and plays a key role in transmission of epigenetic memory across cell divisions and transcription regulation (PubMed:20871596, PubMed:23086925, PubMed:23317504, PubMed:29176719, PubMed:29379197). Remains associated with acetylated chromatin throughout the entire cell cycle and provides epigenetic memory for postmitotic G1 gene transcription by preserving acetylated chromatin status and maintaining high-order chromatin structure (PubMed:22334664, PubMed:23317504, PubMed:23589332). During interphase, plays a key role in regulating the transcription of signal-inducible genes by associating with the P-TEFb complex and recruiting it to promoters (PubMed:16109376, PubMed:16109377, PubMed:19596240, PubMed:23589332, PubMed:24360279). Also recruits P-TEFb complex to distal enhancers, so called anti-pause enhancers in collaboration with JMJD6 (PubMed:16109376, PubMed:16109377, PubMed:19596240, PubMed:23589332, PubMed:24360279). BRD4 and JMJD6 are required to form the transcriptionally active P-TEFb complex by displacing negative regulators such as HEXIM1 and 7SKsnRNA complex from P-TEFb, thereby transforming it into an active form that can then phosphorylate the C-terminal domain (CTD) of RNA polymerase II (PubMed:16109376, PubMed:16109377, PubMed:19596240, PubMed:23589332, PubMed:24360279). Regulates differentiation of naive CD4(+) T-cells into T-helper Th17 by promoting recruitment of P-TEFb to promoters (By similarity). Promotes phosphorylation of 'Ser-2' of the C-terminal domain (CTD) of RNA polymerase II (PubMed:23086925). According to a report, directly acts as an atypical protein kinase and mediates phosphorylation of 'Ser-2' of the C-terminal domain (CTD) of RNA polymerase II; these data however need additional evidences in vivo (PubMed:22509028). In addition to acetylated histones, also recognizes and binds acetylated RELA, leading to further recruitment of the P-TEFb complex and subsequent activation of NF-kappa-B (PubMed:19103749). Also acts as a regulator of p53/TP53-mediated transcription: following phosphorylation by CK2, recruited to p53/TP53 specific target promoters (PubMed:23317504). Isoform B. Acts as a chromatin insulator in the DNA damage response pathway. Inhibits DNA damage response signaling by recruiting the condensin-2 complex to acetylated histones, leading to chromatin structure remodeling, insulating the region from DNA damage response by limiting spreading of histone H2AX/H2A.x phosphorylation.
Alternative names
HUNK1, BRD4, Bromodomain-containing protein 4, Protein HUNK1
Recommended products
Rabbit Polyclonal BRD4 antibody. Suitable for IHC-P, IP, WB and reacts with Mouse, Human samples. Immunogen corresponding to Synthetic Peptide within Human BRD4 aa 1300 to C-terminus.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7 - 8
Preservative: 0.09% Sodium azide
Constituents: Tris citrate/phosphate- Form
- Liquid
- Clonality
- Polyclonal
- Immunogen
- Synthetic Peptide within Human BRD4 aa 1300 to C-terminus. The exact immunogen used to generate this antibody is proprietary information. Database link O60885
- Purification technique
- Affinity purification Immunogen
- Concentration
- Purification notes
ab245285 was affinity purified using an epitope specific to Brd4 immobilized on solid support.
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage duration
- 1-2 weeks
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- -20°C
- Aliquoting information
- Upon delivery aliquot
- Storage information
- Avoid freeze / thaw cycle
Notes
Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.
If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.
Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The Brd4 protein also known as Bromodomain containing protein 4 plays an important role as a transcriptional regulator. It recognizes acetylated lysine residues on histone tails through its bromodomains. This binding impacts chromatin structure and transcriptional activation. The Brd4 protein has a molecular weight of around 150 kDa and is often used as a marker in studies using Brd4 western blot techniques. It mainly expresses in the nucleus of most cell types and is an important focus for developing Brd4 anticuerpos useful for detecting protein size discrepancies in research.
- Biological function summary
Brd4 plays a significant role in regulating gene expression during cell cycle progression. It interacts with the Mediator complex enabling communication between transcriptional activators and RNA polymerase II. This protein is integral for maintaining chromatin in an active state ensuring accessibility for transcription machinery. Moreover Brd4 size and functionality relate to the elongation of transcription by RNA polymerase II ensuring the seamless expression of genes essential for cellular functions.
- Pathways
Brd4 integrates into transcriptional and epigenetic regulation pathways. It acts in the positive regulation of the MYC pathway directly linking Brd4 with proto-oncogene c-Myc. Also Brd4 engages in the JAK/STAT signaling pathway where its activity affects cell growth and immune response. These interactions establish Brd4 as a central regulator within these pathways facilitating communication and coordination among molecular players.
- Associated diseases and disorders
Brd4 link heavily to cancer and inflammatory diseases. Aberrant Brd4 activity often associates with various cancers including leukemia where its regulation impacts oncogenic drivers. Clinically targeting Brd4 presents a therapeutic avenue as its inhibition affects cyclin-dependent kinase 9 (CDK9) an important regulator of the cell cycle. Similarly Brd4 alterations implicate in inflammatory disorders impacting nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) signaling revealing pathways for intervening in pathogenic processes.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
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3 product images
Western blot - Anti-Brd4 antibody (ab245285)
Prepared using NETN lysis buffer.
All lanes: Western blot - Anti-Brd4 antibody (ab245285) at 0.1 µg/mL
Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 15 µg
Lane 2: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 15 µg
Lane 3: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 15 µg
Lane 4: MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate at 15 µg
Lane 5: HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 15 µg
Lane 6: A549 (human lung carcinoma cell line) whole cell lysate at 15 µg
Lane 7: SW620 whole cell lysate at 15 µg
Lane 8: SK-MEL-28 whole cell lysate at 15 µg
Lane 9: Ramos (human Burkitt's lymphoma cell line) whole cell lysate at 15 µg
Developed using the ECL technique.
Predicted band size: 152 kDa
Exposure time: 10s
Immunoprecipitation - Anti-Brd4 antibody (ab245285)
Brd4 was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1 mg for IP, 20% of IP loaded) with ab245285 at 3 μg per reaction. Western blot was performed from the immunoprecipitate using an alternative rabbit anti-Brd4 antibody at a 1/1000 dilution.
Lane 1: ab245285 IP in HeLa whole cell lysate.
Lane 2: Control IgG IP in HeLa whole cell lysate.
Detection: Chemiluminescence with exposure time of 1 second.
Prepared using NETN lysis buffer.
All lanes: Immunoprecipitation - Anti-Brd4 antibody (ab245285)
Predicted band size: 152 kDa
Western blot - Anti-Brd4 antibody (ab245285)
Prepared using NETN lysis buffer.
All lanes: Western blot - Anti-Brd4 antibody (ab245285) at 0.1 µg/mL
All lanes: NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate at 15 µg
Developed using the ECL technique.
Predicted band size: 152 kDa
Exposure time: 3min
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