Rabbit Recombinant Monoclonal BRD4 antibody. Carrier free. Suitable for ICC/IF, WB, IHC-P, Flow Cyt (Intra) and reacts with Mouse, Human, Rat samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: 100% PBS
Liquid
Monoclonal
ICC/IF | IP | ChIP | WB | IHC-P | Flow Cyt (Intra) | |
---|---|---|---|---|---|---|
Human | Tested | Not recommended | Not recommended | Tested | Not recommended | Expected |
Mouse | Tested | Not recommended | Not recommended | Tested | Tested | Tested |
Rat | Expected | Not recommended | Not recommended | Tested | Tested | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Rat, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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. 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:23589332, PubMed:23317504, PubMed:22334664). 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. Also recruits P-TEFb complex to distal enhancers, so called anti-pause enhancers in collaboration with JMJD6. 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:23589332, PubMed:19596240, PubMed:16109377, PubMed:16109376, PubMed:24360279). 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 BActs 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.
Bromodomain-containing protein 4, Protein HUNK1, HUNK1, BRD4
Rabbit Recombinant Monoclonal BRD4 antibody. Carrier free. Suitable for ICC/IF, WB, IHC-P, Flow Cyt (Intra) and reacts with Mouse, Human, Rat samples.
Bromodomain-containing protein 4, Protein HUNK1, HUNK1, BRD4
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: 100% PBS
Liquid
Monoclonal
Yes
EPR25425-23
Affinity purification Protein A
This antibody does not react with Human species for IHC.
Blue Ice
+4°C
ab289893 is the carrier-free version of Anti-Brd4 antibody [EPR25425-23] ab289886
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
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.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
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.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
This data was developed using Anti-Brd4 antibody [EPR25425-23] ab289886, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling Brd4 with Anti-Brd4 antibody [EPR25425-23] ab289886 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nucleus staining was found in rat testis. The section was incubated with Anti-Brd4 antibody [EPR25425-23] ab289886 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
This data was developed using Anti-Brd4 antibody [EPR25425-23] ab289886, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labeling Brd4 with Anti-Brd4 antibody [EPR25425-23] ab289886 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubμle Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
This data was developed using Anti-Brd4 antibody [EPR25425-23] ab289886, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling Brd4 with Anti-Brd4 antibody [EPR25425-23] ab289886 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.
This data was developed using Anti-Brd4 antibody [EPR25425-23] ab289886, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Lysates were prepared from fresh material and used for Western blotting immediately to minimize protein degradation.
Exposure time: 48 seconds.
All lanes: Western blot - Anti-Brd4 antibody [EPR25425-23] (Anti-Brd4 antibody [EPR25425-23] ab289886) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 4: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 152 kDa
Observed band size: 200 kDa
This data was developed using Anti-Brd4 antibody [EPR25425-23] ab289886, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Brd4 with Anti-Brd4 antibody [EPR25425-23] ab289886 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.
This data was developed using Anti-Brd4 antibody [EPR25425-23] ab289886, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Brd4 with Anti-Brd4 antibody [EPR25425-23] ab289886 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
This data was developed using Anti-Brd4 antibody [EPR25425-23] ab289886, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling Brd4 with Anti-Brd4 antibody [EPR25425-23] ab289886 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining was found in mouse testis (PMID:28076398). The section was incubated with Anti-Brd4 antibody [EPR25425-23] ab289886 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
This data was developed using Anti-Brd4 antibody [EPR25425-23] ab289886, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 180 seconds
All lanes: Western blot - Anti-Brd4 antibody [EPR25425-23] (Anti-Brd4 antibody [EPR25425-23] ab289886) at 1/1000 dilution
Lane 1: Mouse testis tissue lysate at 20 µg
Lane 2: Rat testis tissue lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 152 kDa
Observed band size: 200 kDa
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