Recombinant Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7642] to BRD2 - ChIP Grade
- Suitable for: WB, ICC/IF, IHC-P, Flow Cyt (Intra), ChIP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-BRD2 antibody [EPR7642] - ChIP Grade
See all BRD2 primary antibodies -
Description
Rabbit monoclonal [EPR7642] to BRD2 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IHC-P, Flow Cyt (Intra), ChIPmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human BRD2 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P25440 -
Positive control
- WB: HEK293T, Jurkat, MOLT4, NCCIT and HeLa whole cell lysate (ab150035). ICC/IF: HeLa and wild-type HAP1 cells. IHC-P: Human testis tissue. ChIP: Nuclear extract from LNCaP cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20ºC. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7642 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-BRD2 antibody [EPR7642] (ab197865)
- Alexa Fluor® 647 Anti-BRD2 antibody [EPR7642] (ab197866)
- HRP Anti-BRD2 antibody [EPR7642] (ab198536)
- Alexa Fluor® 555 Anti-BRD2 antibody [EPR7642] (ab213347)
- Alexa Fluor® 594 Anti-BRD2 antibody [EPR7642] (ab215507)
- Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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WB | (2) |
1/1000 - 1/10000. Predicted molecular weight: 88 kDa.
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ICC/IF |
Use a concentration of 0.5 µg/ml.
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IHC-P | (2) |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ChIP |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 88 kDa. |
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. |
Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIP
Use at an assay dependent concentration. |
Target
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Function
May play a role in spermatogenesis or folliculogenesis. -
Sequence similarities
Contains 2 bromo domains.
Contains 1 ET domain. -
Domain
One bromodomain is sufficient for a partial interaction with histone H4 acetylated at 'Lys-13'. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 6046 Human
- Omim: 601540 Human
- SwissProt: P25440 Human
- Unigene: 75243 Human
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Alternative names
- BRD 2 antibody
- Brd2 antibody
- BRD2_HUMAN antibody
see all
Images
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All lanes : Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : BRD2 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 88 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab139690 observed at 110 kDa. Red - loading control ab8245 observed at 36 kDa.
ab139690 Anti-BRD2 antibody [EPR7642] was shown to specifically react with BRD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267265 (knockout cell lysate ab257191) was used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. ab139690 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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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.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
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.
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All lanes : Anti-BRD2 antibody [EPR7642] - ChIP Grade (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.
Secondary
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 88 kDa -
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).
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Immunofluorescence analysis of HeLa cells labeling BRD2, with ab139690 at 1/250 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (24)
ab139690 has been referenced in 24 publications.
- Vichaikul S et al. Inhibition of bromodomain extraterminal histone readers alleviates skin fibrosis in experimental models of scleroderma. JCI Insight 7:N/A (2022). PubMed: 35349485
- Deng Y et al. ARV-771 Acts as an Inducer of Cell Cycle Arrest and Apoptosis to Suppress Hepatocellular Carcinoma Progression. Front Pharmacol 13:858901 (2022). PubMed: 35600879
- Chen IP et al. Viral E protein neutralizes BET protein-mediated post-entry antagonism of SARS-CoV-2. Cell Rep 40:111088 (2022). PubMed: 35839775
- Ding D et al. Retinoblastoma protein as an intrinsic BRD4 inhibitor modulates small molecule BET inhibitor sensitivity in cancer. Nat Commun 13:6311 (2022). PubMed: 36274096
- Imaide S et al. Trivalent PROTACs enhance protein degradation via combined avidity and cooperativity. Nat Chem Biol 17:1157-1167 (2021). PubMed: 34675414