Anti-Brd4 antibody (ab75898)
Key features and details
- Rabbit polyclonal to Brd4
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-Brd4 antibody
See all Brd4 primary antibodies -
Description
Rabbit polyclonal to Brd4 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Horse, Dog
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Immunogen
Synthetic peptide corresponding to Human Brd4 aa 1100-1200 conjugated to keyhole limpet haemocyanin.
(Peptide available asab85811) -
Positive control
- WB: HAP1 and HeLa cell lysates; Mouse and rat liver tissue lysates. ICC/IF: MCF7 cells.
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General notes
"Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help."
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75898 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | (1) |
Use a concentration of 5 µg/ml.
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| WB | (1) |
Use a concentration of 1 µg/ml. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa).
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| Notes |
|---|
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ICC/IF
Use a concentration of 5 µg/ml. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa). |
Target
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Function
Plays a role in a process governing chromosomal dynamics during mitosis. -
Tissue specificity
Ubiquitously expressed. -
Involvement in disease
Note=A chromosomal aberration involving BRD4 is found in a rare, aggressive, and lethal carcinoma arising in midline organs of young people. Translocation t(15;19)(q14;p13) with NUT which produces a BRD4-NUT fusion protein. -
Sequence similarities
Contains 2 bromo domains. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 484869 Dog
- Entrez Gene: 23476 Human
- Entrez Gene: 57261 Mouse
- Entrez Gene: 362844 Rat
- Omim: 608749 Human
- SwissProt: O60885 Human
- SwissProt: Q9ESU6 Mouse
- Unigene: 187763 Human
see all -
Alternative names
- Brd4 antibody
- BRD4-NUT FUSION antibody
- BRD4-NUT fusion oncoprotein antibody
see all
Images
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Western blot - Anti-Brd4 antibody (ab75898)All lanes : Anti-Brd4 antibody (ab75898) at 1 µg/ml
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : BRD4 knockout HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 152 kDa
Observed band size: 220 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab75898 observed at 220 kDa. Red - loading control Mouse anti Vinculin observed at 125 kDa.
ab75898 was shown to react with Brd4 in HAP1 wild-type cells in Western blot with loss of signal observed in BRD4 knockout sample. Wild-type HAP1 and BRD4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab75898 and Mouse anti Vinculin overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Western blot - Anti-Brd4 antibody (ab75898)All lanes : Anti-Brd4 antibody (ab75898) at 1 µg/ml
Lane 1 : Liver (Mouse) Tissue Lysate
Lane 2 : Liver (Rat) Tissue Lysate
Lane 3 : Human liver tissue lysate - total protein (ab29889)
Lysates/proteins at 10 µg/ml per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP)
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 152 kDa
Observed band size: 171 kDa why is the actual band size different from the predicted?
Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 150 seconds -
Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody (ab75898)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canadaab75898 (1/500) staining Brd4 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview. -
Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody (ab75898)ICC/IF image of ab75898 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75898, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and HepG2 cells at 5µg/ml.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (27)
ab75898 has been referenced in 27 publications.
- Mukherjee T et al. Epigenetic reader BRD4 supports mycobacterial pathogenesis by co-modulating host lipophagy and angiogenesis. Autophagy 18:391-408 (2022). PubMed: 34074211
- Zhang YY et al. Pretreatment of nucleus pulposus mesenchymal stem cells with appropriate concentration of H2O2 enhances their ability to treat intervertebral disc degeneration. Stem Cell Res Ther 13:340 (2022). PubMed: 35883157
- Fang Y et al. The H3K36me2 methyltransferase NSD1 modulates H3K27ac at active enhancers to safeguard gene expression. Nucleic Acids Res 49:6281-6295 (2021). PubMed: 34107030
- Kanne J et al. Pericentromeric Satellite III transcripts induce etoposide resistance. Cell Death Dis 12:530 (2021). PubMed: 34031359
- Sui S et al. Ferritinophagy is required for the induction of ferroptosis by the bromodomain protein BRD4 inhibitor (+)-JQ1 in cancer cells. Cell Death Dis 10:331 (2019). PubMed: 30988278