Product nameAnti-Brd4 antibody
See all Brd4 primary antibodies
DescriptionRabbit polyclonal to Brd4
Tested applicationsSuitable for: ICC/IF, IP, WBmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Horse, Dog
- This antibody gave a positive signal in Human, Mouse and Rat Liver Tissue Lysates.
"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."
Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
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.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IP||Use at 1 µg/mg of lysate.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa).|
FunctionPlays a role in a process governing chromosomal dynamics during mitosis.
Tissue specificityUbiquitously expressed.
Involvement in diseaseNote=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 similaritiesContains 2 bromo domains.
- Information by UniProt
- Brd4 antibody
- BRD4-NUT FUSION antibody
- BRD4-NUT fusion oncoprotein antibody
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Brd4 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Hu brain whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab75898 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab75898 at 1000µg immunoprecipitating Brd4 in HEK293 whole cell lysate. Sample was incubated with primary antibody (50mM TRIS dilution buffer) and protein G for 16 hours at 4°C. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
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.
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
ab75898 (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.
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.
This product has been referenced in:
- 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). Read more (PubMed: 30988278) »
- 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). Read more (PubMed: 30250532) »