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
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 & Species
See all applications and species dataApplication Species ICC/IF HumanWB MouseHuman
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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: MCF7 cells.
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General notes
"Note: Batches of this product that have a concentration
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as 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
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch 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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB MouseHumanAll applications HorseDogApplication Abreviews Notes ICC/IF (1) Use a concentration of 5 µg/ml.WB (1) Use a concentration of 1 µg/ml. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa).Notes ICC/IF
Use a concentration of 5 µg/ml.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
References (22)
ab75898 has been referenced in 22 publications.
- 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
- Pan M et al. miR-125b-mediated regulation of cell proliferation through the Jagged-1/Notch signaling pathway by inhibiting BRD4 expression in psoriasis. Mol Med Rep 19:5227-5236 (2019). PubMed: 31059052
- Wang C et al. miR-204 enhances p27 mRNA stability by targeting Brd4 in head and neck squamous cell carcinoma. Oncol Lett 16:4179-4184 (2018). PubMed: 30250532
- Chakraborty D et al. A BET Bromodomain Inhibitor Suppresses Adiposity-Associated Malignant Transformation. Cancer Prev Res (Phila) 11:129-142 (2018). PubMed: 29246955
- Wang Y et al. Bromodomain-containing protein 4 is critical for the antiproliferative and pro-apoptotic effects of gambogic acid in anaplastic thyroid cancer. Int J Mol Med 42:161-170 (2018). PubMed: 29717765
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.
-
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.
