Product nameAnti-BRD2 antibody [EPR7642]
See all BRD2 primary antibodies
DescriptionRabbit monoclonal [EPR7642] to BRD2
Tested applicationsSuitable for: WB, Flow Cyt, ICC/IF, IHC-P, ChIPmore details
Unsuitable for: IP
Species reactivityReacts with: Human
Synthetic peptide within Human BRD2 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P25440
- WB: Jurkat, MOLT4, NCCIT and HeLa whole cell lysate (ab150035). ICC/IF: HeLa and wild-type HAP1 cells. IHC-P: Human testis tissue.
Storage instructionsShipped at 4°C. Store at -20ºC.
Storage bufferPreservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
PurityTissue culture supernatant
- Cell Biology
- Proteolysis / Ubiquitin
- Proteasome / Ubiquitin
- Ubiquitin E3 Enzymes
- RING Finger E3 Ligase
- Anti-BRD2 antibody [EPR7642] (Alexa Fluor® 488) (ab197865)
- Anti-BRD2 antibody [EPR7642] (Alexa Fluor® 647) (ab197866)
- Anti-BRD2 antibody [EPR7642] (HRP) (ab198536)
- Anti-BRD2 antibody [EPR7642] (Alexa Fluor® 555) (ab213347)
- Anti-BRD2 antibody [EPR7642] (Alexa Fluor® 594) (ab215507)
- Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
Our Abpromise guarantee covers the use of ab139690 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 88 kDa.|
|Flow Cyt||1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use a concentration of 0.5 µg/ml.|
|IHC-P||1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ChIP||Use at an assay dependent concentration.|
FunctionMay play a role in spermatogenesis or folliculogenesis.
Sequence similaritiesContains 2 bromo domains.
Contains 1 ET domain.
DomainOne bromodomain is sufficient for a partial interaction with histone H4 acetylated at 'Lys-13'.
- Information by UniProt
- BRD 2 antibody
- Brd2 antibody
- BRD2_HUMAN antibody
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: BRD2 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab139690observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab139690 was shown to specifically react with BRD2 when BRD2 knockout samples were used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. ab139690 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
ab139690 staining Brd2 in wild-type HAP1 cells (top panel) and BRD2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab139690 at 0.5μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Overlay histogram showing HeLa cells stained with ab139690 (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 (ab139690, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Chromatin was prepared from LNCaP cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab139690 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
All lanes : Anti-BRD2 antibody [EPR7642] (ab139690) at 1/1000 dilution
Lane 1 : MOLT4 cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : NCCIT cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 88 kDa
Immunofluorescence analysis of HeLa cells labeling BRD2, with ab139690 at 1/250 dilution.
Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling BRD2 with ab139690 at 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This product has been referenced in:
- Chan KH et al. Impact of Target Warhead and Linkage Vector on Inducing Protein Degradation: Comparison of Bromodomain and Extra-Terminal (BET) Degraders Derived from Triazolodiazepine (JQ1) and Tetrahydroquinoline (I-BET726) BET Inhibitor Scaffolds. J Med Chem 61:504-513 (2018). Read more (PubMed: 28595007) »
- Jin X et al. DUB3 Promotes BET Inhibitor Resistance and Cancer Progression by Deubiquitinating BRD4. Mol Cell 71:592-605.e4 (2018). Read more (PubMed: 30057199) »