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AB197608

Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)]

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Anti-Brd4 antibody [EPR5150(2)] - Alexa Fluor® 647 conjugated (ab197608) is a rabbit recombinant monoclonal antibody detecting Brd4 in Flow Cytometry (Intra), ICC/IF. Suitable for Human.

- Biophysical QC for unrivalled batch-batch consistency

View Alternative Names

HUNK1, BRD4, Bromodomain-containing protein 4, Protein HUNK1

4 Images
Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (AB197608)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (AB197608)

ab197608 staining Brd4 in SW480 cells. The cells were fixed with 100% methanol (5 min), permeabilised in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab197608 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 488, shown in green) at 1/167 dilution overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (AB197608)
  • Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (AB197608)

Overlay histogram showing SW480 cells stained with ab197608 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab197608, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 647 used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (AB197608)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (AB197608)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SW480 (human colorectal adenocarcinoma epithelial cell) cells labelling Brd4 with ab197608 at 1/500 compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (AB197608)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (AB197608)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Brd4 with ab197608 at 1/500 compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR5150(2)

Isotype

IgG

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Carrier free

No

Reacts with

Human, Mouse, Human, Mouse

Applications

ICC/IF, Flow Cyt (Intra)

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/50 - 1/500", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/alexa-fluor-647-rabbit-igg-monoclonal-epr25a-isotype-control-ab199093'>ab199093</a> - Rabbit monoclonal IgG (Alexa Fluor®; 647), is suitable for use as an isotype control with this antibody.</p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>" }, "Mouse": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/50 - 1/500", "FlowCytIntra-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>" } } }

Product details

What is this antibody validated in?
Alexa Fluor® 647 Anti-Brd4 antibody [EPR5150(2)] (ab197608) is a rabbit recombinant monoclonal antibody and is validated for use in Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunocytochemistry/immunofluorescence (ICC/IF) in Human samples.

Related products
Antibody clone EPR5150(2) is also available pre-conjugated to a variety of labels for your convenience – Anti-Brd4 Alexa Fluor® 647 [EPR5150(2)] (ab197608).

Other related products
We have a range of other formats of antibody clone [EPR5150(2)] also available for your convenience: ab128874, Carrier free - ab182446, Alexa Fluor® 488 - ab197606, Alexa Fluor® 647 - ab197608, HRP - ab197609, Alexa Fluor® 594 - ab207057

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.4 Preservative: 0.02% Sodium azide Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
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|Store in the dark

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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 protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Target data

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.
See full target information BRD4

Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Frontiers in immunology 16:1586174 PubMed40519909

2025

Small extracellular vesicles from human umbilical cord mesenchymal stem cells delivering miR-202-5p alleviate renal ischemia-reperfusion injury by targeting the GOLIM4/PI3K/AKT axis.

Applications

Unspecified application

Species

Unspecified reactive species

Xiang Peng,Wei Shi,Haitao Yu,Zhenwei Feng,Zongjie Wei,Weiyang He,Xin Gou,Yongpeng Xie
View all publications

Product promise

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